Wheat variety msu line e0028

ABSTRACT

A wheat variety designated MSU Line E0028, the plants and seeds of MSU Line E0028, methods for producing a wheat plant produced by crossing the variety E0028 with another wheat plant, and hybrid wheat seeds and plants produced by crossing the variety E0028 with another wheat line or plant, and the creation of variants by mutagenesis or transformation of variety E0028. The present technology also relates to methods for producing other wheat varieties or breeding lines derived from MSU Line E0028 and to wheat varieties or breeding lines produced by those methods.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/171,982, filed on Apr. 23, 2009. The entire disclosure of the aboveapplication is incorporated herein by reference.

GOVERNMENT RIGHTS

This invention was made with support in part by the U.S. Department ofAgriculture, under Agreement No. 59-0790-6-061, where the invention is acooperative project with the U.S. Wheat & Barley Scab Initiative. TheU.S. Government has certain rights in the invention.

INTRODUCTION

This invention relates to wheat and more particularly to a variety ofsoft winter white wheat designated MSU Line E0028, also referred to as“Ambassador.”

Wheat is grown worldwide and is a widely adapted cereal. Wheat may bedivided into five main market classes, which include the common wheat(Triticum aestivum L.) classes: hard red winter, hard red spring, softred winter, soft and hard white, and durum (Triticum turgidum L.).Common wheats are used in a variety of food products, such as bread,cookies, cakes, crackers, and noodles. In general, the hard wheatclasses are milled into flour used for breads and the soft wheat classesare milled into flour used in a variety of products, such as pastries,crackers, breakfast cereals and soup thickeners. Wheat starch is used inthe food and paper industries, as laundry starches, and in otherproducts.

Grain quality of wheat is very important for its use in baking. To testthe grain quality of wheat for use as flour, milling properties areanalyzed. Important milling properties include relative hardness orsoftness, weight per bushel of wheat (test weight), siftability of theflour, break flour yield, middlings flour yield, total flour yield,flour ash content, and wheat-to-flour protein conversion. Goodprocessing quality for flour is also important. Good qualitycharacteristics for flour from soft wheats include low to medium-lowprotein content, low water absorption, production of large-diameter testcookies and large volume cakes. Wheat glutenins and gliadins, whichtogether confer the properties of elasticity and extensibility, play animportant role in the grain quality. Changes in quality and quantity ofthese proteins change the end product for which the wheat can be used.

Wheat is an important and valuable field crop. Thus, a continuing goalof wheat breeders is to develop stable, high yielding wheat varietiesthat are agronomically sound and have good grain quality for itsintended use. To accomplish this goal, the wheat breeder must select anddevelop wheat plants that have the traits that result in superiorvarieties. These selection processes, which ultimately lead to themarketing and distribution of the wheat variety, can take many yearsfrom the time the first cross is made. Development of new wheatvarieties is therefore a time-consuming process that requires preciseforward planning, efficient use of resources, and a minimum of changesin direction.

SUMMARY

The present technology provides seeds of soft winter white wheat varietydesignated E0028, representative seed of variety E0028 deposited underAmerican Type Culture Collection (ATCC) Patent Deposit DesignationPTA-10223, also denominated “Ambassador.” The present technology alsoprovides compositions and methods that include, use, or operate on, orare derived from E0028. Such technology includes seeds of MSU LineE0028, whole plants and portions of plants of MSU Line E0028, andmethods for producing a wheat plant by crossing MSU Line E0028 withanother wheat plant. Products include flour and other refined orisolated materials derived from variety E0028. For example, theseinclude edible products such as baked goods, cereals, pastas, beverages,livestock feeds, energy products such as biofuels, and further includenon-edible products such as wheat straw and construction materialsproduced from MSU Line E0028.

Methods further include developing other wheat varieties or breedinglines derived from MSU Line E0028 and compositions that include thewheat varieties or breeding lines produced by those methods. Creation ofvariants, by mutagenesis or transformation of MSU Line E0028, is alsoprovided. The present compositions and methods also relate to transgenicbackcross conversions of MSU Line E0028.

MSU Line E0028 demonstrates a unique combination of traits, includinghigher yield (bushels/acre), high flour yield, and increased winterhardiness in comparison to other wheat varieties.

DETAILED DESCRIPTION

The following description of technology is merely exemplary in nature ofthe subject matter, manufacture and use of one or more inventions, andis not intended to limit the scope, application, or uses of any specificinvention claimed in this application or in such other applications asmay be filed claiming priority to this application, or patents issuingtherefrom. The following definitions and non-limiting guidelines must beconsidered in reviewing the description of the technology set forthherein.

The headings (such as “Introduction” and “Summary”) and sub-headingsused herein are intended only for general organization of topics withinthe present technology, and are not intended to limit the disclosure ofthe technology or any aspect thereof. In particular, subject matterdisclosed in the “Introduction” may include novel technology and may notconstitute a recitation of prior art. Subject matter disclosed in the“Summary” is not an exhaustive or complete disclosure of the entirescope of the technology or any embodiments thereof. Classification ordiscussion of a material within a section of this specification ashaving a particular utility is made for convenience, and no inferenceshould be drawn that the material must necessarily or solely function inaccordance with its classification herein when it is used in any givencomposition.

The citation of references herein does not constitute an admission thatthose references are prior art or have any relevance to thepatentability of the technology disclosed herein. All references citedin the “Detailed Description” section of this specification are herebyincorporated by reference in their entirety.

The description and specific examples, while indicating embodiments ofthe technology, are intended for purposes of illustration only and arenot intended to limit the scope of the technology. Moreover, recitationof multiple embodiments having stated features is not intended toexclude other embodiments having additional features, or otherembodiments incorporating different combinations of the stated features.Specific examples are provided for illustrative purposes of how to makeand use the apparatus and systems of this technology and, unlessexplicitly stated otherwise, are not intended to be a representationthat given embodiments of this technology have, or have not, been madeor tested.

As referred to herein, all compositional percentages are by weight ofthe total composition, unless otherwise specified. As used herein, theword “include,” and its variants, is intended to be non-limiting, suchthat recitation of items in a list is not to the exclusion of other likeitems that may also be useful in the materials, compositions, devices,and methods of this technology. Similarly, the terms “can” and “may” andtheir variants are intended to be non-limiting, such that recitationthat an embodiment can or may comprise certain elements or features doesnot exclude other embodiments of the present technology that do notcontain those elements or features.

“A” and “an” as used herein indicate “at least one” of the item ispresent; a plurality of such items may be present, when possible.“About” when applied to values indicates that the calculation or themeasurement allows some slight imprecision in the value (with someapproach to exactness in the value; approximately or reasonably close tothe value; nearly). If, for some reason, the imprecision provided by“about” is not otherwise understood in the art with this ordinarymeaning, then “about” as used herein indicates at least variations thatmay arise from ordinary methods of measuring or using such parameters.In addition, disclosure of ranges includes disclosure of all distinctvalues and further divided ranges within the entire range.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The goal is to combine in a single variety animproved combination of desirable traits from the parental germplasm.These important traits may include higher seed yield, resistance todiseases and insects, tolerance to drought and heat, improved grainquality, and better agronomic qualities.

Field crops are bred through techniques that take advantage of theplant's method of pollination. A plant is self-pollinated if pollen fromone flower is transferred to the same or another flower of the sameplant. A plant is sib-pollinated when individuals within the same familyor line are used for pollination. A plant is cross-pollinated if thepollen comes from a flower on a different plant from a different familyor line. The term cross-pollination herein does not includeself-pollination or sib-pollination. Wheat plants (Triticum aestivumL.), are recognized to be naturally self-pollinated plants which, whilecapable of undergoing cross-pollination, rarely do so in nature. Assuch, intervention for control of pollination is important to theestablishment of superior varieties.

A cross between two different homozygous lines produces a uniformpopulation of hybrid plants that may be heterozygous for many gene loci.A cross of two heterozygous plants each that differ at a number of geneloci will produce a population of plants that differ genetically andwill not be uniform. Regardless of parentage, plants that have beenself-pollinated and selected for type for many generations becomehomozygous at almost all gene loci and produce a uniform population oftrue breeding progeny. A homozygous plant is hereby defined as a plantwith homozygous genes at 95% or more of its loci. The term “inbred” asused herein refers to a homozygous plant or a collection of homozygousplants.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of variety used commercially; e.g., F1 hybrid variety, pure-linevariety, etc. For highly heritable traits, a choice of superiorindividual plants evaluated at a single location can be effective,whereas for traits with low heritability, selection may be based on meanvalues obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.In general, breeding starts with the crossing of two genotypes (a“breeding cross”), each of which may have one or more desirablecharacteristics that is lacking in the other or which complements theother. If the two original parents do not provide all the desiredcharacteristics, other sources can be included by making more crosses.In each successive filial generation, F1→F2; F2→F3; F3→F4; F4→F5, etc.,plants are selfed to increase the homozygosity of the line. Typically ina breeding program five or more generations of selection and selfing arepracticed to obtain a homozygous plant.

Pedigree breeding is commonly used for the improvement ofself-pollinating crops. Two parents that possess favorable,complementary traits are crossed to produce an F1. An F2 population isproduced by selfing or sibbing one or several F1's. Selection of thebest individuals may begin in the F2 population; then, beginning in theF3, the best individuals in the best families are selected. Replicatedtesting of families can begin in the F4 generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F5, F6 and F7), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new varieties.

Backcross breeding is used to transfer genes for simply inherited,qualitative traits from a donor parent into a desirable homozygousvariety that is utilized as the recurrent parent. The source of thetraits to be transferred is called the donor parent. After the initialcross, individuals possessing the desired trait or traits of the donorparent are selected and then repeatedly crossed (backcrossed) to therecurrent parent. The resulting plant is expected to have the attributesof the recurrent parent (e.g., variety) plus the desirable trait ortraits transferred from the donor parent. This approach has been usedextensively for breeding disease resistant varieties.

Each wheat breeding program includes a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulvarieties produced per unit of input; e.g., per year, per dollarexpended, etc.

Various recurrent selection techniques are used to improvequantitatively inherited traits controlled by numerous genes. The use ofrecurrent selection in self-pollinating crops depends on the ease ofpollination and the number of hybrid offspring from each successfulcross. Recurrent selection can be used to improve populations of eitherself- or cross-pollinated crops. A genetically variable population ofheterozygous individuals is either identified or created byintercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. Plantsfrom the populations can be selected and selfed to create new varieties.

Another breeding method is single-seed descent. This procedure, in thestrict sense, refers to planting a segregating population, harvesting asample of one seed per plant, and using the one-seed sample to plant thenext generation. When the population has been advanced from the F2 tothe desired level of inbreeding, the plants from which lines are derivedwill each trace to different F2 individuals. The number of plants in apopulation declines each generation due to failure of some seeds togerminate or some plants to produce at least one seed. As a result, notall of the F2 plants originally sampled in the population will berepresented by a progeny when generation advance is completed. In amultiple-seed procedure, wheat breeders commonly harvest one or morespikes (heads) from each plant in a population and thresh them togetherto form a bulk. Part of the bulk is used to plant the next generationand part is put in reserve. The procedure has been referred to asmodified single-seed descent. The multiple-seed procedure has been usedto save labor at harvest. It is considerably faster to thresh spikeswith a machine than to remove one seed from each by hand for thesingle-seed procedure. The multiple-seed procedure also makes itpossible to plant the same number of seeds of a population eachgeneration of inbreeding. Enough seeds are harvested to make up forthose plants that did not germinate or produce seed.

Bulk breeding can also be used. In the bulk breeding method, an F2population is grown. The seed from the populations is harvested in bulkand a sample of the seed is used to make a planting the next season.This cycle can be repeated several times. In general when individualplants are expected to have a high degree of homozygosity, individualplants are selected, tested, and increased for possible use as avariety.

Molecular markers, including techniques such as Starch GelElectrophoresis, Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs), and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods. One use of molecular markers is Quantitative TraitLoci (QTL) mapping. QTL mapping is the use of markers, which are knownto be closely linked to alleles that have measurable effects on aquantitative trait. Selection in the breeding process is based upon theaccumulation of markers linked to the positive effecting alleles and/orthe elimination of the markers linked to the negative effecting allelesfrom the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against themarkers of the donor parent. Using this procedure can minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program (Openshaw et al.,Marker-assisted Selection in Backcross Breeding. In: ProceedingsSymposium of the Analysis of Molecular Marker Data, 5-6 Aug. 1994, pp.41-43. Crop Science Society of America, Corvallis, Oreg.). The use ofmolecular markers in the selection process may be referred to as MarkerAssisted Selection or as Genetic Marker Enhanced Selection.

The production of double haploids can also be used for the developmentof homozygous lines in the breeding program. Double haploids areproduced by the doubling of a set of chromosomes (1N) from aheterozygous plant to produce a completely homozygous individual. Thiscan be advantageous because the process omits the generations of selfingneeded to obtain a homozygous plant from a heterozygous source. Variousmethodologies of making double haploid plants in wheat have beendeveloped (Laurie, D. A. and S. Reymondie, Plant Breeding, 1991, v.106:182-189; Singh, N. et al., Cereal Research Communications, 2001, v.29:289-296; Redha, A. et al., Plant Cell Tissue and Organ Culture, 2000,v. 63:167-172; and U.S. Pat. No. 6,362,393, Konzak et al., issued Mar.26, 2002.

Though pure-line varieties are the predominate form of wheat grown forcommercial wheat production, hybrid wheat is also used. Hybrid wheatsare produced with the help of cytoplasmic male sterility, nucleargenetic male sterility, or chemicals. Various combinations of thesethree male sterility systems have been used in the production of hybridwheat.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks; e.g., Allard, Principles of Plant Breeding, 1960; Simmonds,Principles of Crop Improvement, 1979; Heyne, Wheat and WheatImprovement, 1987; Allan, “Wheat”, Chapter 18, Principles of CropDevelopment, vol. 2, Fehr editor, 1987).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s). The best lines are candidates for new commercialvarieties; those still deficient in a few traits may be used as parentsto produce new populations for further selection.

A difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior genotype is to observe itsperformance relative to other experimental genotypes and to a widelygrown standard variety. Generally, a single observation is inconclusive,so replicated observations are required to provide a better estimate ofits genetic worth.

A breeder uses various methods to help determine which plants should beselected from the segregating populations and ultimately which lineswill be used for commercialization. In addition to the knowledge of thegermplasm and other skills the breeder uses, a part of the selectionprocess is dependent on experimental design coupled with the use ofstatistical analysis. Experimental design and statistical analysis areused to help determine which plants, which family of plants, and finallywhich lines are significantly better or different for one or more traitsof interest. Experimental design methods are used to control error sothat differences between two lines can be more accurately determined.Statistical analysis includes the calculation of mean values,determination of the statistical significance of the sources ofvariation, and the calculation of the appropriate variance components.Five and one percent significance levels are customarily used todetermine whether a difference that occurs for a given trait is real ordue to the environment or experimental error.

Plant breeding is the genetic manipulation of plants. The goal of wheatbreeding is to develop new, unique, and superior wheat varieties. Inpractical application of a wheat breeding program, the breeder initiallyselects and crosses two or more parental lines, followed by repeatedselfing and selection, producing many new genetic combinations. Thebreeder can theoretically generate billions of different geneticcombinations via crossing, selfing, and mutations. The breeder has nodirect control at the cellular level. Therefore, two breeders will neverdevelop exactly the same line. Each year, the plant breeder selects thegermplasm to advance to the next generation. This germplasm is grownunder unique and different geographical, climatic and soil conditions;and further selections may then be made during and at the end of thegrowing season.

Proper testing should detect major faults and establish the level ofsuperiority or improvement over current varieties. In addition toshowing superior performance, there must be a demand for a new variety.The new variety must be compatible with industry standards, or mustcreate a new market. The introduction of a new variety may incuradditional costs to the seed producer, the grower, processor andconsumer, for special advertising and marketing, altered seed andcommercial production practices, and new product utilization. Thetesting preceding release of a new variety should take intoconsideration research and development costs as well as technicalsuperiority of the final variety. It must also be feasible to produceseed easily and economically.

A wheat variety needs to be highly homogeneous, homozygous, andreproducible to be useful as a commercial variety. There are manyanalytical methods available to determine the homozygotic stability,phenotypic stability, and identity of these varieties. The oldest andmost traditional method of analysis is the observation of phenotypictraits. This type of data is usually collected in field experiments overthe life of the wheat plants to be examined. Phenotypic characteristicsobserved include traits such as seed yield, head configuration, glumeconfiguration, seed configuration, lodging resistance, diseaseresistance, and maturity, among others.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. There are many laboratory-based techniques available forthe analysis, comparison, and characterization of a plant genotype;among these are Gel Electrophoresis, Isozyme Electrophoresis,Restriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asMicrosatellites, and Single Nucleotide Polymorphisms (SNPs). Gelelectrophoresis is particularly useful in wheat. For example, wheatvariety identification is possible through electrophoresis of gliadin,glutenin, albumin and globulin, and total protein extracts (Bietz, J. A.1987, Genetic and biochemical studies of nonenzymatic endospermproteins, In E. G. Heyne (ed.) Wheat and Wheat Improvement, ASA, MadisonWis., pp. 215-241).

The present technology relates to a new and distinctive wheat variety,designated E0028, also referred to as “Ambassador,” which is the resultof years of careful breeding and selection as part of a wheat breedingprogram. Variety E0028, is semi-dwarf soft white winter wheat (Triticumaestivum L.) developed at Michigan State University (MSU). E0028 wasselected from a cross of Pioneer Brand 2737W/D1148 made in 1994 at MSU.The cultivar is an F₇ derived line, and the original experimental numberwith MSU is E0028. In addition to standard yield test criteria, millingand baking performance was also considered for selection. E0028 wasreleased because of its excellent grain yield, flour yield, and goodwinter hardiness. Disadvantages may include low test weight, andsusceptibility to Fusarium head blight (Fusarium graminearum Schwabe)and associated deoxynivalenol (DON) accumulation with respect to otherentries in the Michigan State Performance Trial. E0028 is well adaptedto Michigan and Ontario, Canada, and has also produced high yieldsthroughout the region. The name “Ambassador” was chosen because thecultivar's performance excels in both the U.S. (Michigan) and Canada(Ontario), bringing together white wheat growers on both sides of theborder.

E0028 was selected from the cross Pioneer Brand ‘2737W’(PI#561197)/D1148 made in 1994 at MSU. 2737W is a soft white winterwheat developed by Pioneer Hi-Bred International, Inc. (Johnston, Iowa).D1148 is a soft white winter wheat developed by MSU with the pedigree12214/B0246//‘Yorkstar’ (Jensen, 1968, Registration of ‘Yorkstar’ Wheat(Reg. No. 475). Crop Sci. 8: 641-b-642-b)/3/‘Augusta’ (Everson et al,1986, Registration of ‘Augusta’ wheat. Crop Sci. 26:201-202)/4/‘Asosan’(CI#12665)/4* ‘Genessee’ (CI#12653)//‘Arrow’(Jensen, 1973, Registrationof ‘Arrow’ Wheat. Crop Sci 13(4):495).

E0028 was developed using a modified bulk breeding method. The crossbetween the two parents, designated as cross population 940310, was madein the greenhouse in spring 1994. The Fi was advanced in the greenhousein fall 1994. The F₁ and F₂ were bulk harvested and planted in singledrill plots (7 rows, 15.2 cm spacing, and 2.74 m long) in the field inMason, Mich. in fall 1995 and 1996, respectively. From the F₃ drillplot, F₄ seed was bulk harvested and planted in one space planted fieldplot (15.24 m long, 4 rows, 17.8 cm spacing between seeds within rows)in fall 1997. From the F₄ plot, forty heads were selected (see criteriafor selection below) and planted as F₅ headrows in fall 1998. FourteenF₅ headrows were selected and sampled for three to seven heads, whichwere planted as F₆ headrows in fall 1999. The remainder of the plants inthe selected F₅ headrows was bulk harvested (by headrow) and planted asF₆ drill plots in fall 1999. One F₆ drill plot (entry 155),corresponding with five F₆ headrows (the drill plot and headrows allderived from a single F₅ headrow), was harvested and planted for the2001 Preliminary Yield Trial (PYT) under the experimental linedesignation E0028. Three of the five corresponding F₆ headrows were bulkharvested individually and space-planted as three F₇ plots in fall 2000.From each of the F₇ plots (experimental names E0028-1, E0028-2 andE0028-3), individual plants were harvested and each plant was used tosow a single plant derived space planted F_(7:8) plots (15.24 m long, 4rows, 17.8 cm spacing between seeds within rows) in fall 2001. Theremainder of the E0028-2 F₇ plot was bulk harvested and used to plantthe 2002 Advanced Yield Trial (AYT). One F_(7:8) single plant derivedspace planted plot was selected, from which two plants were harvestedand planted as F_(7:9) space planted plots with the experimental nameE0028-R6. In addition, a bulk of the remainder of the F_(7:8) singleplant derived space planted plot was planted in the 2003 AYT. From thetwo F_(7:9) plots, ten individual plants were selected and planted asF_(7:10) space planted plots. The remainder of plants in the F_(7:9)plots were bulk harvested and planted as F_(7:10) plants in the 2004Michigan State Performance Trial (MSPT). Four of the ten F_(7:10) singleplant derived space planted plots were selected. From these four plots,nineteen plants were harvested and planted as F_(7:11) single plantderived space planted plots planted in fall 2004, and the remainder ofthe plots were bulk harvested and planted as F_(7:11) in the 2005 MSPTunder experimental name E0028-R6. From the nineteen F_(7:11) singleplant derived space planted plots, sixteen were selected and bulkharvested. This bulk was used to plant an F_(7:12) increase andpurification in Colorado in 2005. A bulk of E0028-R6 was used to plantthe F_(7:12) in the 2006 MSPT. In 2006, the seed increase from Coloradowas used to plant the F_(7:13) for a second increase in Colorado, aswell as the 2007 MSPT, Uniform Eastern Soft Red Winter Wheat Nursery(UESRWWN), and OPT. The 2007 Colorado seed increase was used to plantthe 2008 MSPT and OPT.

E0028 (E0028, E0028-2 and E0028-R6) was included in the followingreplicated yield trials harvested in 2001 through 2008: PYT (2001), AYT(2002 and 2003), MSPT (2004 through 2008). Yield trial plots wereplanted in the following manner: All yield trials were planted in sixlocations in various counties around the State of Michigan. Plot sizewas 3.4 m long, 7 rows, 15.24 cm row spacing for 2001 and 2002, but 3.7m long, 7 rows, 15.24 cm row spacing in 2003 and 2004, and 3.7 m long, 6rows, 19 cm row spacing from 2005 onward. Trials were four replicationalpha-lattice designs at each site, except the 2001 PYT, which wasplanted as a three replication alpha-lattice design at each site. Forthe 2001 through 2004 yield trials, seeding rates were the equivalent ofa solid stand of 4.45 million seeds per hectare in 15.24 cm rows. Forthe 2005 yield trial onward, seeding rates were increased to 4.94million seeds per hectare in a solid stand planted in 19.95 cm rows.Fall fertilizer application varied by cooperator practice, and springnitrogen was applied as urea (100.9 kg ha⁻¹ actual N). No foliarfungicides were applied and weeds and insects were controlled as neededwith Harmony™ Extra and Lannate™, respectively. Yield trial plots wereharvested on a single day according to site, except for one location in2008, but analysis of the 2008 data revealed that the different harvestdays did not negatively impact yield or test weight data for that site.For 2003, 2006 and 2008, one, one and two yield trial sites,respectively, were not harvested because of various problems with thosesites. Yield was calculated using the entire area of the plot includingthe wheel tracks between plots, a calculation that tends tounderestimate the total yield. One location was not harvested in 2006due to atypical lodging, and two locations were abandoned in 2008 due tosevere ice and water damage. Yield and test weight data were collectedfor all harvested locations, and all replications of the MSPT in eachyear. Other data were recorded opportunistically, as the traits wereapparent in some years and not others. For each year that trait datawere collected, at least two replications were observed in at least twolocations, with the exception of BYDV, which was observed in only onelocation in 2007. Since grain color is a major distinction betweenvarieties in the trial, grain color was also recorded.

Selection prior to the F₇ was based on visual evaluation of wintersurvival, plant height, grain appearance (including resistance to blackpoint, Alternaria spp. and various fungi), general adult plant reactionto powdery mildew [Blumeria graminis (DC.) E. O. Speer], Wheat spindlestreak mosaic virus (WSSMV), leaf blotches [causal organisms were notspecifically identified, but were likely a combination of Stagonosporanodorum (Berk.) Castellani & E. G. Germano and Septoria tritici Robergein Desmaz], leaf rust [P. recondita Roberge ex Desmaz. f. sp. tritici(Eriks. & E. Henn.) D. M. Henderson], lodging and maturity. Selection ofall of these traits, with the exception of height, grain appearance andmaturity, were very influenced by environmental pressures that could bepresent in some years and not others. Hence, selection for many of thesetraits was opportunistic according to the conditions in each year. Fromthe F₇ onward, E0028 was evaluated in replicated multi-location yieldtrials in Michigan, where milling and baking performance were employedas criteria for selection in addition to standard yield test criteria.

At the seedling stage, E0028 has white/clear (no anthocyanins)coleoptiles, semiprostate juvenile plant growth habit, glabrous withmedium green lower leaf blades and medium tillering capacity at lowdensities. At boot stage, flag leaves are medium green and glabrous(some waxiness on lower side) of medium width and medium length,slightly recurved with green and slightly pubescent auricles, and has aflag leaf sheath that is glabrous with a somewhat waxy bloom. Atmaturity, E0028 is of medium height (90.5 cm, not significantlydifferent from the MSPT trial mean of 89 cm, LSD_(p<0.05)=3.5 cm, Table15), with straight culms with a slightly waxy and glabrous upperinternode, is slightly pubescent on margins of the rachis, and has thickwalled white colored straw. The spike is white, awnletted, inclined,oblong in shape with pronounced waxy bloom and is medium in length anddensity; awns are shorter than the spike and white. Glumes are glabrousand of medium width and length with oblique shoulders and medium lengthobtuse beaks. E0028 has kernels that are soft white, oval to ovate inshape, rounded cheek shape, medium brush with midlong brush hairs.

Variety E0028 is adapted to Michigan, USA and Ontario, Canada. Itdemonstrates a higher yield in bushels per acre and increased winterhardiness versus other wheat varieties tested in the Michigan StatePerformance Trial. E0028 demonstrates good milling and baking qualities,with higher than average flour yield. Softness equivalent percent andpercent protein in flour are average. Lactic acid retention is lowerthan average. E0028 is lower than average for black point (Alternariaspp.).

In comparison with other varieties tested in Michigan, E0028 showsaverage performance to leaf rust (Puccinia recondita f.sp. tritici),powdery mildew (Erysiphe graminis f.sp. tritici), leaf blotch (likely acombination of Stagonospora tritici and Stagonospora nodorum), and wheatspindle streak mosaic virus. Winter kill, plant height, flowering time,and lodging susceptibility are also average.

The primary weaknesses of E0028 include lower than average test weightand susceptibility to Fusarium head blight (Fusarium graminearum) andassociated deoxynivalenol (DON) accumulation. However, Fusarium headblight visual symptoms (incidence, severity, and index) are notstatistically different from average. E0028 also exhibits higher thanaverage pre-harvest sprouting, as compared to other wheat varieties.

E0028 has been found to be uniform and stable in its performance inreplicated yield trials, including over four years of MSPTs. E0028remains essentially unchanged in its primary and distinctivecharacteristics following sexual reproduction. It has self-pollinated asufficient number of generations, with careful attention to uniformityof plant type, to ensure homozygosity and phenotypic stability. The linehas been increased with continued observation for uniformity. Nosubstantial variant traits are observed or are expected in E0028. SinceE0028 is substantially homozygous, it may be reproduced by plantingseeds of the line, growing the resulting wheat plants underself-pollinating or sib-pollinating conditions, and harvesting theresulting seed, using techniques familiar to the agricultural arts.Variants, though infrequent, have been observed as follows: brown chaff<0.03%, extreme talls <0.05%, bearded <0.06%, red seed <0.5%. Thesevariants are at commercially acceptable levels.

The yield of E0028 is slightly better over a 4 year average, though notsignificant, than the white wheat variety known as “Calcdonia.”Calcdonia (Reg. no. CV-943, PI 610188) is a soft white winter wheat(Triticum aestivum L.) developed by the Cornell Agricultural ExperimentStation (Cornell Univ., Ithaca, N.Y.) that exhibits high grain yield andwide-adaptation in the northeastern USA and southern Ontario, Canada.Variety E0028 exhibits good winter hardiness in comparison to Calcdoniaand grows about 2 inches taller than Calcdonia.

Selection characteristics for E0028 include higher yield (bushels/acre),higher flour yield, and an increase in winter hardiness, in comparisonto averages of other wheat varieties, including D8006W, Crystal, Jewel,Aubrey, Pioneer Brand 25W41, and Calcdonia. However, E0028 exhibitslower than average test weight and higher than average DON in the grain,as compared to averages of the other wheat varieties.

The early harvest test weight of variety E0028 can be ascertained todetermine whether the test weight of E0028 is improved by earlierharvesting. The following method is used to examine the early harvesttest weight. Sub-samples of 3 feet (out of 12 foot plots) from qualitytrials at 5 yield testing sites are manually harvested. A singlereplication is harvested at each of the five locations. E0028 isharvested, as well as four other varieties for comparison. These samplesare harvested between 2 and 6 days earlier than the regular harvest forthe site. Early harvested samples are threshed and run through amoisture and test weight machine the same day as harvest, then placed ondriers at Michigan State University's wheat barn.

The remainder of the quality trial plots (9 feet) is used for earlyharvest, plus the quality trial plots at the 6th yield testing site (12feet—not used for early harvest) are harvested at the regular harvestingtime using a combine. These samples are also dried on the drier. Driedsamples of the early harvest and the regular harvest from the qualitytrial plots are sent to Star of the West Milling (Frankenmuth, Mich.)for testing of falling number and DON levels.

Characteristics of MSU Line E0028 are summarized and categorizedaccording to the several data types presented in the Tables below.Several entries are included for comparison, including MSU releases (MSUD8006W, MSU E0027, MSU E1007W) in addition to widely grown varieties,such as Calcdonia. All entries are white wheat. Where applicable,significant differences in comparison with E0028 are denoted as “S+” fora value significantly more than E0028 (LSD 0.05), or as “S−” for a valuesignificantly less than E0028 (LSD 0.05). Breeder interpretation isneeded to consider whether higher (S+) or lower (S−) values for a traitare desirable.

TABLE 1 E0028 Early Harvest Test Weight TEST % FALLING WEIGHTS MOISTUREPPM DON NUMBER Normal Early Normal Early Normal Early Normal EarlyHarvest Harvest Harvest Harvest Harvest Harvest Harvest Harvest Name2007 2007 2007 2007 2007 2007 2007 2007 MSU Line E0028 57.3 57.6 12.416.1 0.11 0.20 311 317 Crystal (MSU E0027) 58.0 56.4 12.1 18.1 0.27 0.38329 324 AC Mountain 57.3 54.8 12.8 18.8 0.17 0.32 334 340 Aubrey 59.5 S+58.7 13.2 S+ 16.1 <0.1 0.11 351 326 Caledonia 57.4 54.8 12.9 18.4 0.440.48 330 334 MEANS 58.4 56.1 13.4 17.8 0.31 0.30 338 333 LSD (0.05)  0.9—  0.8 — — — — — CV (%)  1.4 —  5.1 — — — — — S+ = significantly morethan E0028 (LSD 0.05) S− = significantly less than E0028 (LSD 0.05) DON= deoxynivalenol

TABLE 2 Source of Samples relating to Early Harvest Test Weight inTable 1. Trait Regular Harvest Early Harvest Test Weight StatePerformance Trial Plots Subsample of quality trial (4 reps, 6 sites)plots (1 rep, 5 sites) Moisture State Performance Trial Plots Subsampleof quality trial (4 reps, 6 sites) plots (1 rep, 5 sites) DON (ppm)Remainder of quality trial Subsample of quality trial plots (1 rep, 6sites) plots (1 rep, 5 sites) Falling Remainder of quality trialSubsample of quality trial Number plots (1 rep, 6 sites) plots (1 rep, 5sites)

Tables 3 through 13 present analysis of data from the normal harvesttimes for the State Performance Trial and the Fusarium Head Blight (FHB)screening nursery. These trials are performed by the Department of Cropand Soil Sciences, College of Agriculture and Natural Resources atMichigan State University, East Lansing, Mich. Data from the WheatPerformance Trials is categorized by year and is available online at[www.css.msu.edu/varietytrials/wheat/Variety_Results.html].

TABLE 3 Single and Multiyear Harvest Data Averages - Data from MSU WheatState Performance Trials (4 reps per site, see parentheses for thenumber of sites each year). Test Weight: lbs/Bushel Single Year Data: 4reps/site Multi-Year Averages (6 sites) (5 sites) (6 sites) (6 sites) 2YR 3 YR 4 YR Name 2007 2006 2005 2004 06-07 05-07 04-07 MSU Line E002857.3 56.4 57.3 53.6 56.9 57.0 56.2 MSU D8006W 58.1 56.0 57.8 53.7 57.157.3 56.4 Crystal (MSU E0027) 58.0 56.1 57.4 55.2 S+ 57.1 57.2 56.7Jewel (MSU E1007W) 59.1 S+ 57.3 59.3 S+ 56.1 S+ 58.2 S+ 58.6 S+ 58.0 S+Aubrey 59.5 S+ 58.9 S+ 59.8 S+ 58.4 S+ 59.2 S+ 59.4 S+ 59.2 S+ PioneerBrand 25W41 59.2 S+ 57.4 S+ 60.0 S+ 56.7 S+ 58.3 S+ 58.9 S+ 58.3 S+Caledonia 57.4 56.3 58.7 S+ 54.8 56.9 57.5 56.8 Trial Mean 58.4 57.458.9 56.7 58.2 58.5 58.1 LSD (0.05)  0.9  1.0  0.8  1.3  1.0  1.0  1.1CV (%)  1.4  1.3  1.3  2.0  0.9  1.0  1.4 S+ = significantly more thanE0028 (LSD 0.05) S− = significantly less than E0028 (LSD 0.05)

TABLE 4 Single and Multiyear Harvest Data Averages - Data from MSU WheatState Performance Trials (4 reps per site, see parentheses for thenumber of sites each year). Yield: Bushels/Acre (Adjusted to 13%Moisture) Single Year Data: 4 reps/site Multi-Year Averages (6 sites) (5sites) (6 sites) (6 sites) 2 YR 3 YR 4 YR Name 2007 2006 2005 2004 06-0705-07 04-07 MSU Line E0028 94.0 99.5 82.2 72.0 96.8 91.9 86.9 MSU D8006W94.6 97.9 80.8 74.6 96.3 91.1 87.0 Crystal (MSU E0027) 93.3 97.1 82.269.4 95.2 90.9 85.5 Jewel (MSU E1007W) 91.3 94.6 80.6 75.5 93.0 88.885.5 Aubrey 87.9 S− 87.8 83.2 75.1 87.9 S− 86.3 S− 83.5 Pioneer Brand25W41 88.3 S− 93.4 79.1 69.9 90.9 S− 86.9 82.7 Caledonia 83.1 S− 94.879.6 70.2 89.0 S− 85.8 S− 81.9 Trial Mean 87.7 90.9 80.0 72.8 90.3 87.584.1 LSD (0.05)  4.1 7.0 4.3 4.6  5.1  5.4 5.7 CV (%)  4.1 6.2 4.7 5.6 2.8  3.8 4.8 S+ = significantly more than E0028 (LSD 0.05) S− =significantly less than E0028 (LSD 0.05)

TABLE 5 Single and Multiyear Harvest Data Averages - Samples collectedfrom inoculated FHB screening trial and analyzed via ELISA at MSU unlessotherwise noted. DON (ppm) in grain Multi-Year Single Year DataAverages** 2006 2 YR 3 YR Name 2006 2005 2004 GCMS * 05-06 04-06 MSULine  9.0  4.0 13.5  5.6 6.5  8.8 E0028 MSU D8006W  5.6 S− 10.5 S+ 14.5 5.2 8.1 10.2 Crystal (MSU  5.5 S−  8.6 S+ 10.0 S−  4.5 7.1  8.0 E0027)Jewel (MSU  9.3  8.0 14.0  6.5 8.7 10.4 E1007W) Aubrey  4.2 S−  4.8  9.0S−  3.0 S− 4.5  6.0 S− Pioneer Brand  4.8 S−  9.0 S+ 10.5  3.1 S− 6.9 8.1 25W41 Caledonia  4.5 S−  5.3 10.5  3.3 S− 4.9  6.8 Trial Mean  3.1 3.1  7.2  2.3 3.3  4.6 LSD (0.05)  2.0  4.1  3.3  1.5 2.8  2.4 CV (%)32.1 58.3 23.2 31.9 40.7 31.3 S+ = significantly more than E0028 (LSD0.05) S− = significantly less than E0028 (LSD 0.05)

TABLE 6 Single and Multiyear Harvest Data Averages - Field observationsin inoculated FHB Screening Nursery unless otherwise indicated. FHBIncidence (% of spikes) Multi-Year Averages Single Year Data 2 YR 3 YRName 2007 2006 2005 2004* 06-07 05-07 MSU Line E0028 59.3 60.0 57.7 90.059.7 59.0 MSU D8006W 62.8 60.0 80.0 80.0 61.4 67.6 Crystal (MSU E0027)43.2 35.0 S− 83.8 79.0 39.1 S− 54.0 Jewel (MSU E1007W) 70.7 70.0 62.975.0 S− 70.4 67.9 Aubrey 59.1 50.0 64.4 86.0 54.6 57.8 Pioneer Brand25W41 66.1 40.0 77.5 83.0 53.1 61.2 Caledonia 56.3 50.0 60.7 82.0 53.255.7 Trial Mean 54.7 41.5 66.5 82.0 49.2 55.2 LSD (0.05) 21.8 21.7 22.114.0 16.3 17.0 CV (%) 20.5 26.2 20.8  8.0 16.4 18.9 *In 2004 data werecombined between the inoculated trial and a naturally infected site. S+= significantly more than E0028 (LSD 0.05) S− = significantly less thanE0028 (LSD 0.05)

TABLE 7 Single and Multiyear Harvest Data Averages - Field observationsin inoculated FHB Screening Nursery unless otherwise indicated. FHBSeverity (% within spikes) Multi-Year Averages Single Year Data 2 YR 3YR Name 2007 2006 2005 2004* 06-07 05-07 MSU Line E0028 45.8 60.0 37.883.0 52.9 47.9 MSU D8006W 52.6 50.0 50.4 74.0 51.3 51.0 Crystal (MSUE0027) 67.3 S+ 40.0 S− 58.0 60.0 S− 53.7 55.1 Jewel (MSU E1007W) 57.945.0 43.8 60.0 S− 51.5 48.9 Aubrey 35.5 45.0 29.2 76.0 40.3 36.6 PioneerBrand 25W41 36.3 40.0 S− 56.5 61.0 S− 38.2 44.3 Caledonia 55.8 65.0 60.375.0 60.4 60.4 Trial Mean 43.6 41.2 36.8 69.0 42.1 42.3 LSD (0.05) 21.016.4 24.0 16.0 15.9 13.7 CV (%) 24.2 20.0 40.1 11.3 18.7 19.8 *In 2004data were combined between the inoculated trial and a naturally infectedsite. S+ = significantly more than E0028 (LSD 0.05) S− = significantlyless than E0028 (LSD 0.05)

TABLE 8 Single and Multiyear Harvest Data Averages - Field observationsin inoculated FHB Screening Nursery unless otherwise indicated. FHBIndex (% overall infection) Multi-Year Averages Single Year Data 2 YR 3YR Name 2007 2006 2005 2004* 06-07 05-07 MSU Line E0028 27.2 36.0 23.568.8 31.6 28.9 MSU D8006W 35.6 30.0 39.6 50.7 32.8 35.1 Crystal (MSUE0027) 30.8 14.0 S− 47.6 44.4 22.4 30.8 Jewel (MSU E1007W) 41.0 31.524.5 52.1 36.3 32.3 Aubrey 22.3 23.0 17.3 19.2 22.7 20.9 Pioneer Brand25W41 24.5 16.0 S− 43.4 59.2 20.3 28.0 Caledonia 33.0 37.0 36.3 68.335.0 35.4 Trial Mean 25.3 18.1 25.0 50.5 21.9 24.2 LSD (0.05) 19.9 16.018.7 29.6 12.7 12.0 CV (%) 39.6 44.0 45.7 27.9 28.6 30.4 *In 2004 FHBindex data was calculated from a naturally infected site. S+ =significantly more than E0028 (LSD 0.05) S− = significantly less thanE0028 (LSD 0.05)

TABLE 9 Multiyear Harvest Data Averages - Data from MSU Wheat StatePerformance Trials. Milling and Baking Properties (04-06) PercentPercent Lactic Acid Flour Yield Protein In Flour Retention Multi-YearMulti-Year Multi-Year Averages Averages Averages 2 YR 3 YR 2 YR 3 YR 2YR 3 YR Name 05-06 04-06 05-06 04-06 05-06 04-06 MSU Line E0028 73.373.1 7.8 7.5  93.6  97.9 MSU D8006W 73.3 73.2 8.2 7.6 112.7 S+ 114.5 S+Crystal (MSU E0027) 72.8 72.4 7.7 7.4 101.0 104.4 S+ Jewel (MSU E1007W)72.0 S− 71.8 S− 8.0 7.4 106.7 S+ 108.8 S+ Aubrey 71.7 S− 72.2 S− 8.5 S+8.1 S+ 103.6 S+ 104.7 S+ Pioneer Brand 25W41 70.9 S− 70.9 S− 7.6 7.4 94.2  94.8 Caledonia 72.4 S− 72.3 7.9 7.5  97.5  98.4 Trial Mean 71.471.2 8.0 7.6 102.4 103.9 LSD (0.05)  0.9  0.9 0.7 0.5  7.9  4.9 CV (%) 0.6  0.7 4.0 4.0  3.8  2.8 S+ = significantly more than E0028 (LSD0.05) S− = significantly less than E0028 (LSD 0.05)

TABLE 10 Multiyear Harvest Data Averages - Data from MSU Wheat StatePerformance Trials. Milling and Baking Properties (04-06) SoftnessEquivalent Quality Lab Test Percent Weight Multi-Year Multi-Year BlackPoint % Averages Averages Multi-Year 2 YR 3 YR 2 YR 3 YR 2006 2 YR Name05-06 04-06 05-06 04-06 HARVEST 05-06 MSU Line E0028 57.9 57.9 61.2 61.1 2.5  6.6 MSU D8006W 59.7 59.5 61.2 61.2 26.1 S+ 25.8 S+ Crystal (MSUE0027) 59.0 58.0 61.3 61.5  1.2  3.4 Jewel (MSU E1007W) 57.2 57.3 61.861.8  3.2  5.6 Aubrey 58.4 58.7 62.9 S+ 62.9 S+  7.5  9.1 Pioneer Brand25W41 63.0 S+ 62.1 S+ 62.3 S+ 62.3 S+ 16.9 S+ 17.5 S+ Caledonia 60.260.4 61.5 61.4  5.1  8.4 Trial Mean 58.6 58.6 62.4 62.4 10.4 12.6 LSD(0.05)  3.2  2.6  1.1  0.8  9.2  6.0 CV (%)  2.7  2.7  0.9  0.8 54.823.3 S+ = significantly more than E0028 (LSD 0.05) S− = significantlyless than E0028 (LSD 0.05)

TABLE 11 Multiyear Harvest Data Averages - Data from MSU Wheat StatePerformance Trials (years included are indicated). Percent Lodging GrainScore Flowering Plant Moisture at (0-9); Date (Days Height Harvest (0 =none) Past Jan. 1) (Inches) 4 YR 3 YR 3 YR 3 YR Grain Chaff Name 04-0704-06 05-07 05-07 Color Color Awns MSU Line E0028 13.3 3.5 152.0 36.3WHITE WHITE NO MSU D8006W 13.4 4.0 152.2 36.5 WHITE WHITE YES Crystal(MSU E0027) 13.0 2.5 153.0 S+ 35.4 WHITE WHITE YES Jewel (MSU E1007W)14.1 S+ 3.0 152.2 36.4 WHITE WHITE YES Aubrey 14.8 S+ 2.7 151.7 36.6WHITE WHITE NO Pioneer Brand 25W41 14.1 S+ 3.6 152.2 34.3 S− WHITE WHITEYES Caledonia 13.9 S+ 3.6 152.7 34.3 S− WHITE WHITE NO Trial Mean 14.44.1 152.2 35.8 LSD (0.05)  0.5 1.9  0.8  1.3 CV (%)  2.6 28.6   1.0  2.2S+ = significantly more than E0028 (LSD 0.05) S− = significantly lessthan E0028 (LSD 0.05)

TABLE 12 Multiyear Harvest Data Averages - Data from MSU Wheat StatePerformance Trials (years included are indicated). Leaf Powdery Leaf InHead Rust Mildew Blotch Sprouting Score Score Score Score (0-9) (0-9)(0-9) (0-9) 3 YR 3 YR 3 YR 3 YR Name 05-07 05-07 04-06 05-07 MSU LineE0028  3.8  2.8 4.1  8.7 MSU D8006W  3.2  1.8 3.9  7.3 Crystal (MSUE0027)  2.6  1.9 4.2  8.4 Jewel (MSU E1007W)  2.9  3.1 3.4  8.3 Aubrey 3.0  1.3 S− 3.5  8.5 Pioneer Brand 25W41  2.2 S−  5.4 S+ 3.5  6.7 S−Caledonia  4.2  3.5 3.9  8.7 Trial Mean (2007 = 67  3.1  2.6 3.7  5.9Entries) LSD (0.05)  1.4  1.2 1.0  1.7 CV (%) 28.2 26.8 16.1 17.8 S+ =significantly more than E0028 (LSD 0.05) S− = significantly less thanE0028 (LSD 0.05)

TABLE 13 Single year data from MSU Wheat State Performance Trials (yearsincluded are indicated). Barley Wheat Yellow Spindle Winter Dwarf StreakKill Stripe Rust Virus Mosaic (Injury) Score Score Virus Score Score(0-9) (0-9) (0-9) (0-9) Name 2007 2007 2006 HARVEST 2005 MSU Line E0028 1.7  4.0 2.0  0.8 MSU D8006W  0.7  0.6 S− 1.0  1.1 Crystal (MSU  5.0 S+ 1.0 S− 2.0  0.6 E0027) Jewel (MSU  2.0  0.6 S− 1.0  1.2 E1007W) Aubrey 0.3  1.8 S− 3.0  1.7 Pioneer Brand  0.0 S−  1.9 S− 1.0  1.9 25W41Caledonia  1.0  2.3 S− 2.0  4.5 S+ Trial Mean  0.9  2.1 3.3  1.8 LSD(0.05)  1.5  1.7 3.1  1.6 CV (%) 103.0 56.2 45.3 55.5 S+ = significantlymore than E0028 (LSD 0.05) S− = significantly less than E0028 (LSD 0.05)

The following characteristics of variety E0028 are ascertained bybreeder evaluation of the data presented in Tables 1-13. Variety E0028performs well in terms of yield and flour yield. It also performsacceptably for milling and baking qualities. It is not significantlydifferent from average for multiyear data for other traits, includingflowering time, lodging, black point, leaf rust, leaf blotch, andpowdery mildew.

The deoxynivalenol (DON) levels of E0028 are higher than average, thoughnot significantly different from other wheat varieties, nor are theysignificantly different from Calcdonia, which is widely grown inMichigan. With respect to the DON levels, there are white wheatvarieties in commercial production with significantly lower levels ofDON. Exceeding acceptable DON levels at the elevator may result in theinability of the farmer to sell the grain. In some cases, growers may bepenalized for higher levels of DON, up to a point, and after this pointthe elevator will not purchase the grain. Shown in Table 14 below, arethe U.S. federal advisory limits for DON for various final products.

TABLE 14 U.S. Federal Advisory Limits for Deoxynivalenol (DON) in partsper million (ppm). DON ppm Type of Products 1 Finished wheat productsfor human consumption. 5 Grain and grain byproducts destined for swineand other animal species (except cattle and chickens); not to exceed 20percent of the diet for swine, and not to exceed 40 percent for otheranimal species. 10 Grain and grain byproducts for ruminating beef andfeedlot cattle older.

Varying levels of controlling Fusarium Head Blight (FHB) anddeoxynivalenol (DON) levels may be possible through fungicideapplications. However, consideration should also be given to the factthat years of data show red wheat varieties in Michigan, in general,have lower DON levels than white wheat varieties in Michigan. It is notyet known if this is a genetic factor in red versus white wheat. Itshould be noted that the DON levels of the red wheat varieties arefactored into the mean of the State Performance Trial (the mean datareported here), thereby decreasing the overall wheat average relative tothe score for E0028. FHB visual symptoms (% incidence, % severity, and %FHB Index) are also high in E0028, but they are not significantlydifferent from the average performance of varieties in the StatePerformance Trial when multiyear data are considered. Fusarium damagedkernels, a trait sometimes considered by breeding programs, has not beeninvestigated with respect to the performance of E0028 for thischaracteristic.

Test weight of E0028 is lower than average, and although the data do notprovide a definitive statistical comparison, it appears that earlierharvest would likely not significantly increase the test weight.However, despite the test weight being low, it is not significantly lessthan the Calcdonia variety and two other soft white wheat varieties fromMichigan State University.

Experimental details for the Michigan State Wheat Performance Trials anddata collection for MSU Line E0028 include the following aspects. Wheatplots are 12 feet long and have 6 rows at 7.5″ row spacing. The trial isdesigned and executed as a four replication alpha-lattice (14 blocks of5 plots each) at all sites except the scab screening nursery. All seedis treated, but the chemicals and rates of use vary according to thepreferences of the originating organization. Seeding rates per linearfoot of row are standardized to the rate that would equate with a standof 2.0 million seeds per acre in a solid stand planted in 7.5″ rows.Fall fertilizer application varies with cooperator practice. Springnitrogen is applied as urea (90 lbs/acre actual N) at green-up. Nofoliar fungicides are applied at any site. Weeds are chemicallycontrolled as needed. All plots at a site are harvested on a single day.Yield is calculated using the entire area of the plot including thewheel tracks between plots. This approach tends to underestimate yield.Data reported as scores is based on a 0-9 scale, where 0 is the bestpossible score. Yield, test weight, and grain moisture data are acquiredelectronically on the plot combine at the time of harvest. Yield data isstandardized to 13% moisture.

Data are collected for flowering date, leaf rust, stripe rust, plantheight, powdery mildew, barley yellow dwarf virus, and leaf blotch. Theflowering date indicates the average number of days past January 1stthat a given entry reached the point where one-half of its heads areflowering. Leaf rust, stripe rust, powdery mildew, barley yellow dwarfvirus, and leaf blotch scores are recorded as “0=no visual symptoms ofdisease present”. Plant height is reported as the distance in inchesfrom the ground to the tip of average heads in a plot. Leaf and striperust scores are based on infection observations of primarily the flagleaf. Powdery mildew scores are based on observations of the entireplant including the flag leaf. Barley Yellow Dwarf Virus (BYDV) istransmitted through aphids and is enhanced with cool temperatures andrain. Barley Yellow Dwarf scores may not be reflective of actualresistance because some cultivars may have seed treated withinsecticides prior to planting. Early infestations (fall) of aphids maybe controlled using certain seed treatments, masking BYDVsusceptibility. The causal organism(s) of the leaf blotching are notidentified, but are likely a combination of Stagonospora tritici,(formerly known as Septoria tritici), and S. nordorum.

Data collection for wheat spindle streak mosaic virus, sprouting, andFusarium head blight include the following details. Wheat spindle streakmosaic virus is transmitted into wheat roots via a soil borne funguscalled Polymyxa graminis. Infections take place during cool, wetperiods. The optimal temperature for symptoms to appear is between 48°F. and 55° F. Sprouting data is based on a greenhouse evaluation of 5heads. Heads are collected within four hours of harvest and dried forapproximately seven days. Scores are taken after the heads are subjectedto near-continuous misting for three to four days. A score of zeroindicates that sprouting was not present. A score of 9 indicates manyshoots and roots observed in the heads during scoring.

Data on Fusarium head blight (scab) are obtained from the Inghammisted/inoculated scab screening nursery. The Ingham scab nursery isinoculated (from lab-produced infected grain spread onto the field), andartificial misting is employed throughout the entire flowering period.Each wheat head (i.e., ‘spike’) is comprised of roughly 14-22“spikelets,” which bear the developing seed. Spikelets that prematurelydie because of scab infection are called “scabby” spikelets. Fieldsymptom data are based on: 1) the percent of spikes showing any scabbyspikelets; 2) the percent of scabby spikelets within infected spikes;and 3) the percent of scabby spikelets considering all spikes (scabindex). The scab index is a measure of the extent of damage to entireplots due to scab infection, and generally relates to the effect of scabon yield. Deoxynivalenol (DON) data is from harvested grain in theinoculated, mist irrigated, scab screening nursery. DON data ispresented in parts per million (ppm). The grain was analyzed at MichiganState University using an ELISA kit (Veratox® for DON5/5, Product #8331)from Neogen® (Lansing, Mich.) in 2005, and at the University ofMinnesota using gas chromatography mass spectrometry from 2006-2008(Fuentes, R. G., Mickelson, H. R., Busch, R. H., Dill-Macky, R., Evans,C. K., Thompson, W. G., Wiersma, J. V., Xie, W., Dong, Y., and Anderson,J. A. 2005, Resource Allocation and Cultivar Stability in Breeding forFusarium Head Blight Resistance in Spring Wheat. Crop Sci. 45:1965-1972).

Black point is the discoloration of the embryo (germ) end andsurrounding areas of the wheat kernel. The embryo tip shows a black tobrown discoloration that may extend into the crease of the kernel.Visual observations consist of 500 seed lots from one rep at each of twolocations observed. Data includes the average percent of kernelsdiscolored. The milling and baking quality data are determined by theUSDA Eastern Soft Wheat Quality Laboratory in Wooster, Ohio. Flour yieldis the ratio of the weight of extractable flour to the weight of milledgrain, expressed as a percentage. Lactic Acid Retention is used by somesoft wheat processors as a measure of protein strength. Higher “softnessequivalent percents” indicate a softer grained wheat.

Traits measured in replicated trials conducted from 2001 onward inMichigan included grain yield (kg ha⁻¹, adjusted to 13% moisture), testweight (kg hL⁻¹), flowering date (days past January 1), plant height(cm) and black point (percent observed post-harvest). Additionally, thefollowing traits were observed and recorded on a 0 to 9 scale, where 0was desirable; plant lodging, winter injury, leaf rust, stripe rust(Puccinia striiformis Westend.), powdery mildew, leaf blotch Barleyyellow dwarf virus (BYDV), and Wheat spindle streak mosaic virus(WSSMV).

In each year that E0028 was included in a Michigan yield trial(2001-2008), it was also included in MSU's artificially inoculated andoverhead irrigated FHB screening nursery. Plot sizes, inoculationmethods and number of replications evaluated in the FHB nursery variedby year. The FHB data from the MSPT reported here (2006-2008) were fromtrials inoculated with wheat grain, plus barley grain in 2007, colonizedwith Fusarium graminearum lineage 7. Inoculum was spread in the trial ata rate of 27 kg ha⁻¹ to 33.6 kg ha⁻¹ (rate varied by year) perinoculation date. Inoculum was applied twice, approximately two weeksapart, to the entire trial. The first application date was applied whenit was predicted that anthesis would approximately two to three weekslater. Genotypes were planted as 1 row plots, 4 feet long in 2006 and 5feet long in 2007 and 2008. Overhead (mist) irrigation was employed onan hourly schedule following the application of the inoculum and throughthe completion of flowering. Data on both incidence and severity werecollected and used to calculate the FHB index (FHBindex=incidence*severity/100) to represent the percentage of overall FHBinfection in a plot. DON mycotoxin data were also collected from samplesharvested from the FHB nursery field trials. DON from the 2006 through2008 harvests was quantified in parts per million using gaschromatography mass spectrometry at the University of Minnesota (Fuenteset al., 2005, Resource Allocation and Cultivar Stability in Breeding forFusarium Head Blight Resistance in Spring Wheat. Crop Sci. 45:1965-1972).

E0028 was also evaluated at additional sites through cooperativeregional tests, including the 2007 Uniform Eastern Soft Red Winter WheatNursery (UESRWWN, coordinated by USDA-ARS, 59 cooperators in 22 USstates) and the 2007 and 2008 OPT (10 sites, Ontario, Canada). Traitdata collected in the UESRWWN included a wide range of data pertainingto agronomics, disease and pest resistance, quality, and molecularmarker data for a select group of loci. Data collected in the OPT alsoincluded agronomic traits and disease resistance.

From 2005 through 2008, a grain sample of 400 to 500 g of each cultivarentered in the MSPT was obtained from a 1:1 mix of two yield-triallocation harvests in Michigan. Sites used for the mixture each year wereselected for absence of pre-harvest sprouting and a minimum of othergrain defects such as Fusarium damaged kernels. Samples from yield trialsites in the following counties were mixed as follows for each year:Saginaw and Midland Counties in 2005, 2007 and 2008; Saginaw and LenaweeCounties in 2006. Prior to milling, samples were aspirated to removebroken, shrunken, and diseased kernels. Whole grain moisturedetermination used low speed corrugated rolls to coarsely break grainand then the oven drying method (Method 44-15A) of the AmericanAssociation of Cereal Chemistry (AACC Approved Methods, 10^(th) ed.,2000), to measure grain moisture content. Samples were then tempered to15% grain moisture. Tempered grain samples were milled after 48 hours toallow for equal water distribution throughout the kernel. Samples weremilled using modifications to AACC Method 26-50 as described by Finneyand Andrews (1986) Revised microtesting for soft wheat qualityevaluation. Cereal Chem. 63:177-182. Milling of 200 g samples wasconducted on a modified Quadrumat Junior flour mill in a controlledtemperature and humidity room (19 to 21° C. and RH 55% to 60%). Productfrom the mill was recovered for sifting on a Great Western Sifter Box toseparate mill product into bran (particles remaining above a 470 μmmesh), flour mids (between 470 and 180 μm mesh) and fine flour(particles passing through a 180 μm mesh). Flour mids were furtherprocessed through reduction milling with smooth rolls on a secondQuadrumat Junior mill and sieved on a Great Western Sifter box using a213 μm mesh screen to produce baking quality flour. Softness equivalent(SE) was calculated as fine flour (flour passing through the 180 μm meshscreen) recovered from the first milling process expressed as a percentof the total weight of milled grain. Softness equivalent is an estimatorof break flour yield from a long-flow multi-stream flour mill. Flouryield was corrected based on softness equivalent to predict flour yieldon the long-flow experimental Allis Chalmers mill at the Soft WheatQuality Laboratory (Gaines et. al 2000, Developing Agreement betweenVery Short Flow and Longer Flow Mills. Cereal Chemistry 77: 187-192).

Flour quality was analyzed using the solvent retention capacity (SRC)test, AACC Method 56-11 (AACC 2000), with the following modifications.Flour sample size for the SRC analysis was 1 g, suspended in 5 g ofsolvent. Disposable glass centrifuge tubes were used for the analysis.Suspension of the flour in the solvent used a vortex mixer for 20 s at 5min intervals for 15 min. All other protocols conformed to the AACCstandard method. Lactic acid SRC was used for all four years of theevaluations; all other solvents were used for flour analysis only in2007 and 2008. Initial flour moistures were determined by oven method(AACC Method 44-16). Flour protein was determined using Near Infra-RedReflectance (SpectraStar 2400, Unity Scientific, Columbia Md.)calibrated by combustion analysis (rapidN III, Elementar Instruments,Cambridge UK) of a subset of samples each year. Baking evaluation forsoft wheat quality used the micro sugar-snap cookie method (AACC Method10-52). The diameters across two cookies were measured at four differentpositions of the cookies, and the four measurements were averaged andreported as the sum of two cookie diameters. A visual assessment oftopgrain cracking of the cookie surface also was scored using a 0 to 9scale by comparing to photographs of standard cookies. Greater amountsof cracking of the cookie surface indicate greater collapse of thecookie at the end of the baking sequence and are generally preferred forsoft wheat quality. Greater score values indicate greater amounts ofsurface cracking.

MSU yield trials (PYT, AYT, MSPT) and associated FHB screening trialswere planted in replicated designs, either alpha-lattice designs orrandomized complete block designs. Data from these trials were analyzedby residual maximum likelihood (Patterson and Thompson, 1972, “Recoveryof inter-block information when block sizes are unequal” Biometrika 58:545-554) in the program “REML” (Thompson et al., 1982, REML a Programfor the Analysis of Non Orthogonal Data by Restricted MaximumLikelihood, COMPSTAT: Proceedings in Computational Statistics. PhysicaVerlag, Wien, 231-232), which not only takes into account locations andreplications, but neighboring plots as well. The coefficients ofvariations and the Least Significant Differences (LSDs) using a P-valueof 0.05 were also determined. For quality analyses, an F-test forsignificance of genotypes used an analysis of variance in SAS PROC GLM(SAS version 9.0, 2003) assuming that genotypes were fixed effects andyears were random effects and that the error term for genotypes was theinteraction of genotypes and years. When possible, trait data arepresented as averages across years (three-year averages except foryield, test weight and quality, for which four year averages arepresented), though not all traits were assessed in all years.

The majority of data presented below were collected on the MSPT. Forease of comparisons between varieties, only data of entries present inall years from 2005 to 2008 are included in Tables 15 through 19. Trialmeans, LSDs and CVs reported in Tables 15 to 18 were determined usingdata of the entire trial each year, of which the reported cultivars hereare only a subset.

In MSPT trials, the three year average of the flowering date of E0028(152.3 Julian Days Flowering) is the same as the trial mean (Table 15).In addition, lodging (2.8) and winter injury (0.8) are not significantlydifferent from the two year and one year trial means for these traits(89 cm, 3.5, 1.8, respectively, LSD_(p<0.05) for lodging and winterinjury =1.8 and 1.6, respectively).

TABLE 15 Agronomic traits of E0028 (i.e., “Ambassador”) in comparisonwith other wheat varieties included in the MSPT. The scale used forevaluation of each trait is shown in parenthesis after the name of thetrait. Flowering Date, Winter days past 1 Height Lodging Injury Januarycm 0-9 0-9 3 YR† 3 YR 2 YR 1 YR Cultivar Cultivar Reference Class 06-08‡06-08 05-06 2005 Ambassador§ This publication SWW 152.3 90.5 2.8 0.825R47 PVP¶ 200200232, 2003 SRW 152.1 83.3 3.3 1.3 25W41 PVP 200300328,2004 SWW 152.4 86.0 3.9 1.9 AC Mountain NR#, Agriculture Canada SWW152.9 99.8 3.9 0.8 Aubrey Private Company†† SWW 151.7 90.0 2.1 1.7 BravoPVP 200000326, 2002 SRW 150.9 94.8 3.0 1.1 Caledonia Sorrells et al.,2004 SWW 152.7 86.0 2.4 4.5 Coral NR, MSU SWW 153.6 95.3 3.8 1.1 CrystalPVP 200800367 SWW 153.3 88.0 2.7 0.6 D6234 PVP 200300259, 2004 SWW 153.193.0 3.6 2.2 D8006 PVP 200500308, 2006 SWW 152.0 90.8 4.0 1.1 DF101Private Company SRW 151.2 87.5 3.2 1.0 Emmit NR, Hyland Seeds SRW 153.090.3 2.9 na‡‡ Hopewell Campbell et al., 2001 SRW 152.4 91.8 2.0 1.3Jewel PVP 200700408, 2008 SWW 152.4 91.0 2.5 1.2 McCormick Griffey etal., 2005a SRW 151.6 78.5 5.1 2.3 MCIA Oasis NR, Ohio State UniversitySRW 152.2 98.5 3.1 na R045 Private Company SRW 151.9 87.0 4.5 0.8 R055Private Company SRW 152.0 84.8 2.3 1.8 Red Amber NR, MSU SRW 152.9 93.33.6 2.0 Red Ruby PVP 200700409 SRW 153.1 89.3 2.5 1.1 Roane Griffey etal., 2001 SRW 151.6 83.0 3.9 1.1 Tribute Griffey et al., 2005b SRW 151.282.8 4.2 1.6 Trial Mean 152.3 89.0 3.5 1.8 LSD_(p<0.05) 0.7 3.5 1.8 1.6CV (%) 0.8 2.4 25.5 55.5  †The number of years averaged is shown by“YR”, with the details of the years included below. ‡Years that areaveraged are indicated by the last two digits of the first and lastyears of the range of years included (e.g. 06-08 = average of 2006, 2007and 2008). §Ambassador (i.e., E0028) listed first, with other entriessorted by name. ¶Plant Variety Protection (PVP) number given #NR = NotRegistered. Culativars listed as NR have known origin, and these originsare indicated. ††Cultivars listed as “Private Company” were provided byprivate companies without information of their origin. ‡‡na, notavailable

The four-year (2005 through 2008) MSPT average yield of E0028 (6160 kgha⁻¹) was significantly greater than the four-year trial mean (5931 kgha⁻¹, LSD_(p<0.05)=242) (Table 16). However, when examining each yearseparately, the yield of E0028 was only significantly greater than thetrial means in 2006 and 2007. Of the varieties presented in Table 16,only ‘25R47’ was consistently higher yielding than E0028 across in eachof the four years, though this difference was only significant in 2005(25R47, 6005 kg ha⁻¹; E0028 5528 kg ha⁻¹; LSD_(p<0.05)=289 kg ha⁻¹), andthe four year average yield (6348 kg ha⁻¹) was not significantlydifferent. In comparison with ‘Calcdonia’, the most widely grown softwhite wheat in Michigan, E0028's yield was consistently greater in eachof the four years, and was significantly greater for the four-yearaverage (Calcdonia=5723 kg ha⁻¹). Regarding test weight, E0028'sfouryear average (74.1 kg hL⁻¹) was significantly less than the trialmean (76.1 kg hL⁻¹, LSD_(p<0.05)=0.9) as well as all other cultivarsincluded in Table 16 with the exceptions of ‘Crystal’ (74.6 kg hL⁻¹) and‘AC Mountain’ (74.6 kg hL⁻¹).

TABLE 16 Yield (kg ha⁻¹) adjusted to 13% moisture, and test weight (kghL⁻¹) for MSPT entries. Four year averages for yield and test weight areshown, in addition to the yield of each individual year. Yield Test kgha⁻¹ (Adjusted to 13% Moisture) weight 4 YR† 1 YR kg hL⁻¹ Cultivar Class05-08‡ 2008 2007 2006 2005 4 YR 05-08 25R47§ SRW 6348 6254 6395 67456005 75.3 Ambassador SWW 6160 6100 6322 6691 5528 74.1 Red Ruby SRW 61606147 6234 6631 5629 77.2 MCIA Oasis SRW 6106 6066 6335 6530 5501 75.3Hopewell SRW 6100 6133 6221 6328 5710 75.9 D8006 SWW 6100 6026 6362 65845434 75.0 Crystal SWW 6093 6032 6274 6530 5528 74.6 R045 SRW 6079 63486328 6322 5319 76.4 Emmit SRW 6053 6026 6079 6449 5649 76.1 R055 SRW6012 5609 6120 6631 5696 76.3 Red Amber SRW 5979 5945 6160 6409 539375.9 DF101 SRW 5972 5844 6032 6369 5642 77.5 Coral SWW 5965 5864 60466422 5521 75.9 Jewel SWW 5952 5891 6140 6362 5420 76.2 Tribute SRW 58985528 6248 6322 5488 78.8 25W41 SWW 5857 5878 5938 6281 5319 76.6 D6234SWW 5824 5958 5844 6100 5400 76.6 AC Mountain SWW 5817 5831 5750 63155367 74.6 Aubrey SWW 5804 5810 5911 5905 5595 77.2 Caledonia SWW 57235575 5588 6375 5353 75.1 Bravo SRW 5696 5723 5555 5925 5568 76.8 RoaneSRW 5521 5548 5676 5676 5178 78.4 McCormick SRW 5266 5192 5118 5535 521277.7 Trial Mean 5931 5871 5898 6113 5380 76.2 LSD_(p<0.05) 242 309 276471 289 0.9 CV (%) 2.9 3.8 4.1 6.2 4.7 0.8 †The number of years averagedis shown by “YR”, with the details of the years included below. ‡Yearsthat are averaged are indicated by the last two digits of the first andlast years of the range of years included (e.g. 06-08 = average of 2006,2007 and 2008). §Entries are sorted by highest 4 year average yield.

Data from the UESRWWN [www.ars.usda.gov/main/docs.htm?docid=2925] andthe OPT [gocereals.ca/] further supported the yield test weightperformance that was observed in Michigan. For the UESRWWN yield data,which included forty-four entries evaluated at twenty-two sites, E0028had an average yield of 5248 kg ha⁻¹, which was greater than the trialmean (4791 kg ha⁻¹) as well a the four checks ‘Foster’ (4529 kg ha⁻¹),Tatton'(4629 kg ha⁻¹), ‘Roane’ (5140 kg ha⁻¹) and ‘INW0411’ (4603 kgha⁻¹). For test weight, which was evaluated at twenty-one sites, E0028had an average test weight of 72.9 kg hL⁻¹, which was lower than themean (75.0 kg hL⁻¹), as well as the four checks (Foster, 74.8 kg hL⁻¹;Patton, 74.3 kg hL⁻¹; Roane, 76.6 kg hL⁻¹; INW0411, 73.8 kg hL⁻¹). TheOPT data are reported according to Areas, which are defined according toclimatic conditions. Area 1 is the southwest portion of Ontario, whichis the warmest, Area II is less warm and has more snowfall, and Area IIIis eastern Ontario, which has more ice accumulation. Among eight softwhite winter wheat cultivars, E0028 showed the highest yield (5622 kgha⁻¹), on a two-year average for Areas I and II combined (mean=4354 kgha⁻¹, 12 sites included), and Area III alone for a two-year average(E0028 yield=5622 kg ha⁻¹, mean=5406 kg ha⁻¹, 6 sites included). E0028'stest weight, however, was the lowest for its class in each of Areas I(73.3 kg 0.50, II and III. Additional data can be viewed at the 2008winter wheat report at http://gocereals.ca/.

E0028 has been characterized for disease and insect resistance inMichigan and through cooperative regional evaluations (Tables 17 and18). Considering the three-year averages (2006 through 2008) of FHBincidence, severity and index, only the percent FHB severity (55.9%) wassignificantly greater than the three-year average of the MSPT (41.0%,LSD_(p<0.05)=14.4) (Table 17). Although E0028's FHB index (28.4%) wasnot significantly different from the three-year average (26.1%,LSD_(p<0.05)=15.6), amongst the other cultivars in Table 17 it is onlyless than Calcdonia (42.0%), the most widely grown soft white wheat inMichigan. FHB incidence for E0028 (69.9%) was not significantlydifferent from the three year average of the MSPT (62.3%,LSD_(p<0.05)=16.7). Knott et al., (2008, Comparison of selection methodsfor the development of white-seeded lines from red×white soft winterwheat crosses. Crop Sci 48: 1807-1816) showed that although visualsymptoms of red vs. white grained wheat are not significantly different,the associated mycotoxin accumulation in white wheat is higher than redwheat. In comparison with other white wheat cultivars included in theMSPTs, the three-year average of E0028 for DON (10.0 ppm) is higher thanall other cultivars listed in table 17 with the exception of Crystal(10.4 ppm), and is significantly higher than ‘Aubrey’ (4.1 ppm,LSD_(p<0.05)=5.6).

TABLE 17 Fusarium head blight (FHB) disease symptoms and deoxynivalenol(DON) mycotoxin accumulation shown in parts per million (ppm) for MSPTentries. Severity Incidence % within Index % of infected % overall DONspikes spikes infection ppm† 3 YR‡ Cultivar Class 06-08§ Ambassador¶ SWW69.9 55.9 40.0 10.0 25R47 SRW 71.9 39.2 28.4 4.6 25W41 SWW 66.5 38.325.4 7.3 AC Mountain SWW 52.1 50.8 26.8 4.9 Aubrey SWW 63.5 35.0 21.84.1 Bravo SRW 69.9 39.8 28.6 4.0 Caledonia SWW 66.9 59.9 42.0 8.3 CoralSWW 51.6 36.8 20.8 5.3 Crystal SWW 59.2 51.8 30.2 10.4 D6234 SWW 67.051.0 32.3 6.0 D8006 SWW 72.3 51.0 37.8 7.9 DF101 SRW 68.9 31.8 21.2 3.8Emmit SRW 51.5 42.1 21.4 3.6 Hopewell SRW 71.6 46.1 33.8 5.3 Jewel SWW74.9 46.1 34.6 9.6 McCormick SRW 57.6 28.5 14.3 2.5 MCIA Oasis SRW 60.942.8 26.6 4.2 R045 SRW 61.8 35.6 22.0 4.9 R055 SRW 60.4 38.3 22.6 3.5Red Amber SRW 60.9 53.1 32.3 6.5 Red Ruby SRW 63.0 43.3 31.0 6.3 RoaneSRW 61.0 32.4 19.3 2.6 Tribute SRW 50.7 30.7 15.1 3.6 Trial Mean 62.341.0 26.1 3.8 LSD_(p<0.05) 16.7 14.4 15.6 5.6 CV (%) 16.3 21.5 36.6 42.0†ppm = parts per million ‡The number of years averaged is shown by “YR”,with the details of the years included below. §Years that are averagedare indicated by the last two digits of the first and last years of therange of years included (e.g. 06-08 = average of 2006, 2007 and 2008).¶Ambassador (i.e. E0028) listed first, other entries sorted by name.

TABLE 18 Disease response to Leaf and Stripe Rust, Powdery Mildew (PM),Barley Yellow Dwarf Virus (BYDV) and Wheat Spindle Streak Mosaic Virus(WSSMV) for MSPT entries. Quantification of the % of black pointobserved on harvested grain. Leaf Stripe Black Rust Rust PM BYDV WSSMVPoint 0-9 % 3 YR‡ 1 YR 3 YR 1 YR 1 YR 3 YR Cultivar Class 06-08§ 200706-08 2007 2006 05-07 Am- SWW 5.2 1.7 2.8 4.0 2 11.7 bassador 25R47 SRW2.9 0 3.7 0.6 3 21.7 25W41 SWW 3.3 0 4.6 1.9 1 37.5 AC SWW 4.9 1.0 3.32.9 2 18.3 Mountain Aubrey SWW 4.5 0.3 1.6 1.8 3 10.0 Bravo SRW 5.8 05.0 3.2 2 11.3 Caledonia SWW 5.1 1.0 3.8 2.3 2 12.1 Coral SWW 3.7 3.74.4 2.1 1 14.5 Crystal SWW 3.6 5.0 2.0 1.0 2 3.1 D6234 SWW 3.3 3.3 2.70.7 1 40.2 D8006 SWW 4.4 0.7 2.1 0.6 1 27.5 DF101 SRW 3.5 0.3 1.2 2.2 712.5 Emmit SRW 4.6 4.3 3.8 4.5 6 36.3 Hopewell SRW 5.2 0 3.2 1.5 1 5.0Jewel SWW 4.0 2.0 3.5 0.6 1 8.4 Mc- SRW 6.7 0 0.5 1.0 2 20.9 CormickMCIA SRW 0.8 0.7 1.5 2.9 1 na§ Oasis R045 SRW 3.2 3.0 3.4 1.1 3 34.3R055 SRW 3.5 0.7 2.6 1.1 4 34.2 Red SRW 3.3 0 1.3 1.0 2 20.0 Amber RedRuby SRW 3.8 3.0 2.9 0.4 1 11.7 Roane SRW 3.2 0.3 3.4 0.8 9 5.3 TributeSRW 0.2 2.7 0.3 1.6 4 31.3 Trial 3.8 0.9 2.7 2.1 3.3 17.6 MeanLSD_(p<0.05) 1.5 1.5 1.1 1.7 3.1 15.2 CV (%) 23.7 103 24.5 56.2 45.3 53†The number of years averaged is shown by “YR”, with the details of theyears included below. ‡Years that are averaged are indicated by the lasttwo digits of the first and last years of the range o years included(e.g. 06-08 = average of 2006, 2007 and 2008). §na = not applicable

E0028's responses to additional diseases are shown in Table 18. E0028 issusceptible to leaf rust. Although E0028's susceptibility (5.2) to leafrust was not significantly worse than the three-year trial mean (3.8,LSD_(p<0.05)=1.5), only ‘McCormick’ (6.7) and ‘Bravo’ (5.8) had higherleaf rust values. The susceptibility of ‘Hopewell’ (5.2), the mostwidely grown soft red wheat in Michigan, was equivalent to E0028. Forpowdery mildew, E0028 (2.8) was not significantly different from thetrial mean (2.7, LSD_(p<0.05)=1.1). However, BYDV data from 2007 showthat E0028 (4.0) is significantly worse than the trial mean (2.1,LSD_(p<0.05)=1.7), being surpassed only by ‘Emmit’ (4.5) forsusceptibility. In 2006, WSSMV was present and rated in Michigan, andE0028 was more resistant (2.9) than the trial mean (3.3), beingsignificantly less susceptible than Roane, Emmit and ‘DF101’ (9.0, 6.0,and 7.0, respectively, LSD_(p<0.05)=3.1). For post-harvest evaluationsof percent black point, E0028 (11.7) was more resistant than the trialmean (17.6), and was significantly better than many other varieties(LSD_(p<0.05)=15.2).

Stem and leaf rust were evaluated with multiple races through theUESRWWN at the USDA Cereal Disease Rust Laboratory, Minn. For stem rust,seedling reactions of E0028 to Puccinia graminis f. sp. tritici (P.graminis Pers.:Pers. f. sp. tritici Eriks. & E. Henn.) U.S. races QFCS,QTHJ, RCRS, RKQQ, TPMK, TTTT, TTKS were susceptible, though there was alow infection frequency for TPMK and TTTT, and mostly zero infection forRKQQ (heterogeneous reaction). Based on reactions to leaf rust racesBBBD, MFPS, MJBJ, MCRK, KFBJ, MHDS, TGBG, TNRJ, it is postulated thatE0028 has gene Lr9 relating to disease resistance.

E0028 has good soft wheat quality for pastry products based on millingand flour measurements. Calcdonia was the most widely grown soft whitewinter wheat in the eastern U.S. at the time of E0028's release. It iswell accepted in the industry as a cultivar with targeted soft whitequality attributes. E0028 had greater flour yield than Calcdonia atsimilar levels of softness equivalent and flour protein concentration(Table 19). Based on four years of evaluations, E0028 and Calcdonia havesimilar sugar snap two-cookie mean diameters (18.86 and 18.65 cm,respectively, LSD_(p<0.05)=0.51). Top grain scores (0 to 9 scale, where9 is desirable) for E0028 and Calcdonia also were similar (4.3 and 4.9,respectively, LSD_(p<0.05)=1.5). The solvent retention capacity profilefor E0028 was not significantly different from Calcdonia for any of thefour solvents evaluated (Table 19). E0028 also had favorable milling andbaking quality by comparison to the standard public soft red wintercultivar, Hopewell. E0028 had significantly greater flour yield thanHopewell with similar softness equivalent. E0028 had a greatersugar-snap two-cookie diameter than Hopewell (18.86 and 18.33 cm,respectively, LSD_(p<0.05)=0.51). Overall water absorption as measuredby the SRC method was less for E0028's flour than for Hopewell's flour(50.33 and 51.83 g 100 g⁻¹, respectively, LSD_(p<0.05)=1.39). This wasreflected in the lower sodium carbonate SRC for E0028 than Hopewell(64.38 and 70.18 g 100 g⁻¹, respectively, LSD_(p<0.05)=2.85), whichsuggests that the E0028 flour samples have less damaged starch due tomilling than do the Hopewell samples. Based on the lactic acid SRC,E0028 has less gluten strength than Hopewell. E0028 had a lactic acidSRC of 96.68 g 100 g⁻¹ and Hopewell 111.55 g 100 g⁻¹.

Analysis of E0028 in 2008 and 2009 by the USDA-ARS Regional Small GrainsGenotyping Laboratory, Raleigh N. C., showed that E0028 has theoverexpressing allele for Bx7oe, contributing towards gluten strength(Glu-B1al). However, since the other high molecular weight glutenins areweak, notably the Glu-D1a allele at the Glu-D1 locus, E0028 is onlymoderate for gluten strength compared with other eastern soft winterwheat genotypes.

TABLE 19 Milling and baking quality for E0028 soft white winter wheat,Michigan trials, 2005 to 2008†. Sugar-snap cookie Solvent retentioncapacity solvents Flour Softness Flour Top grain Sodium Lactic CultivarClass yield equivalent protein Diameter‡ score Water§ carbonate§Sucrose§ acid g 100 g⁻¹ cm 0-9¶ g 100 g⁻¹ Ambassador SWW 73.1 58.1 7.6118.86 4.3 50.33 64.36 84.53 87.25 25R47 SRW 72.6 62.6 7.36 19.14 5.150.55 65.25 84.71 95.43 25W41 SWW 70.4 62.3 7.47 18.74 4.8 52.70 67.8388.85 89.49 AC Mountain SWW 71.6 58.0 7.68 18.51 5.0 51.21 65.53 84.5586.21 Aubrey SWW 71.3 59.6 8.20 17.99 3.7 52.14 66.76 87.39 102.16 BravoSRW 70.4 54.8 8.37 18.30 3.8 52.18 67.23 90.25 89.36 Caledonia SWW 72.158.4 7.91 18.65 4.9 50.58 65.80 83.69 96.95 Coral SWW 72.0 59.2 7.6618.73 4.8 50.13 65.09 84.25 95.10 Crystal SWW 72.6 57.9 7.61 18.57 4.551.62 66.33 84.25 91.65 D6234 SWW 70.1 55.3 8.09 18.37 4.2 52.71 68.5084.70 78.46 D8006 SWW 73.3 60.5 7.96 18.67 4.6 51.22 65.87 86.67 107.41DF101 SRW 68.9 50.8 8.56 17.68 4.3 54.19 68.03 94.40 113.42 Emmit SRW72.5 57.0 7.72 18.28 4.3 53.11 67.52 87.60 79.52 Envoy# SWW 71.9 53.98.17 18.22 4.0 54.21 68.58 90.55 105.74 Hopewell SRW 69.4 60.0 7.9818.33 4.3 51.83 70.18 88.55 109.69 Jewel SWW 71.5 56.6 7.89 17.97 4.253.32 70.10 90.35 101.17 McCormick SRW 70.2 59.0 8.52 17.76 4.8 55.3571.93 94.95 108.54 MCIA Oasis SRW 72.1 58.1 7.85 18.69 4.8 51.12 65.4885.80 100.75 Pearl†† SWW 71.0 59.0 8.04 18.41 3.7 52.56 69.58 88.60106.38 R045 SRW 72.2 59.2 7.81 18.46 4.5 54.66 71.21 89.70 91.45 R055SRW 72.3 58.8 7.86 18.52 5.8 52.09 64.00 87.55 94.20 Red Amber SRW 71.454.7 8.41 18.42 4.3 50.76 66.13 85.95 103.95 Red Ruby SRW 71.0 60.9 7.6918.56 4.3 51.89 68.24 89.75 103.61 Roane SRW 68.5 58.0 8.06 17.46 3.056.26 73.16 98.70 107.52 Tribute SRW 70.2 53.3 8.19 17.92 4.0 57.5573.38 97.45 111.78 Average 71.3 57.7 7.96 18.37 4.4 52.65 67.95 88.6998.85 LSD_(p<0.05) 0.8 3.0 0.48 0.43 1.3 0.97 1.91 3.47 7.27 F-test forCultivar 24.9*** 8.13*** 3.89*** 8.36*** 1.58 16.44*** 8.06*** 5.94***16.17*** *F-test of cultivar variances significant at the 95% CI whentested with cultivar x year variance as the denominator. ***F-test ofcultivar variances significant at the 99.9% CI when tested with cultivarx year variance as the denominator. †Trials produced in Michigan andevaluated by the USDA-ARS Soft Wheat Quality Laboratory. ‡Sum of twocookie diameters. §Water, sucrose, and sodium carbonate SRC evaluationswere only conducted in 2007 and 2008. ¶For the top grain score, 9 isdesirable. #Envoy was included in the 2005 AYT, and the MSPT following.It was licensed exclusively from MSU in 2008. ††Although Pearl wasincluded in the MSPT from 2005-2008, not all data was reported.Therefore, only the quality data is being reported for these years (PVP#200300114, 2003).

The present technology further includes the following aspects withrespect to MSU Line E0028.

Reproduction of the MSU Line E0028 can occur by tissue culture andregeneration. Tissue culture of various tissues of wheat andregeneration of plants therefrom are performed using well known andwidely published methods. A review of various wheat tissue cultureprotocols can be found in Maheshwari et al. (1995), In vitro culture ofwheat and genetic transformation: Retrospect and Prospect, CriticalReviews in Plant Science, 14:149-178. Thus, another aspect of thepresent technology is to provide cells or tissue, which upon growth anddifferentiation, produce wheat plants capable of having thephysiological and morphological characteristics of MSU Line E0028.

As used herein, a wheat plant includes various portions of the wheatplant such as plant protoplasts, plant cell tissue cultures from whichwheat plants can be regenerated, plant calli, plant clumps, and plantcells that are intact in plants or portions of plants, such as embryos,pollen, ovules, pericarp, seed, flowers, florets, heads, spikes, leaves,roots, root tips, anthers, and the like.

Molecular biological techniques allow the isolation and characterizationof genetic elements with specific functions, such as those encodingspecific protein products. The genome of plants can be engineered tocontain and express foreign genetic elements, or additional, or modifiedversions of native or endogenous genetic elements, in order to alter thetraits of a plant in a specific manner. Any DNA sequence, whether from adifferent species or from the same species, which is inserted into thegenome by transformation, is referred to herein collectively as atransgene. Several methods for producing transgenic plants have beendeveloped, and embodiments of the present technology relate totransformed versions of the MSU Line E0028.

Numerous methods for plant transformation exist, including biologicaland physical, plant transformation protocols. See, for example, Miki BL, Fobert P, Charest P J, Iyer V N, (1993) Procedures for introducingforeign DNA into plants, In: Glick B R, Thompson J E, eds., Methods inplant molecular biology and biotechnology, Boca Raton, USA: CRC Press,67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber M Y, Crosby W L (1993) Vectors forplant transformation, In B R Glick, J E Thompson, Eds., Methods in PlantMolecular Biology and Biotechnology, CRC Press, Baton Rouge, La., pp89-119.

The most prevalent types of plant transformation involve theconstruction of an expression vector. Such a vector comprises a DNAsequence that contains a gene under the control of or operatively linkedto a regulatory element, for example a promoter. The vector may containone or more genes and one or more regulatory elements.

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include but are not limited to genes;coding sequences; inducible, constitutive, and tissue specificpromoters; enhancing sequences; and signal and targeting sequences.

A genetic trait, engineered into a wheat plant with transformationtechniques, can be moved or crossed into another line using traditionalbreeding techniques that are well known in the plant breeding arts. Forexample, a backcrossing approach could be used to move a transgene froma transformed wheat plant to an elite wheat variety and the resultingprogeny would comprise a transgene. As used herein, “crossing” can referto a simple X by Y cross, or the process of backcrossing, depending onthe context. The term “breeding cross” excludes the processes of selfingor sibbing.

With transgenic plants according to the present technology, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney G. and Orr G. A., The purificationof avidin and its derivatives on 2-iminobiotin-6-aminohexyl-Sepharose4B, Anal. Biochem. 114(1): 92-6 (1981).

According to one embodiment, the transgenic plant provided forcommercial production of foreign protein is a wheat plant. In anotherembodiment, the biomass of interest is seed. A genetic map can begenerated, primarily via conventional RFLP, PCR, and SSR analysis, whichidentifies the approximate chromosomal location of the integrated DNAmolecule. For exemplary methodologies in this regard, see Glick B R,Thompson J E, editors, Methods in plant molecular biology andbiotechnology, (CRC Press, Boca Raton, 1993). Map information concerningchromosomal location is useful for proprietary protection of a subjecttransgenic plant. If unauthorized propagation is undertaken and crossesmade with other germplasm, the map of the integration region can becompared to similar maps for suspect plants, to determine if the latterhave a common parentage with the subject plant. Map comparisons wouldinvolve hybridizations, RFLP, PCR, SSR and sequencing, all of which areconventional techniques.

Likewise, by means of the present technology, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Through the transformation of wheat the expression of genescan be modulated to enhance disease resistance, insect resistance,herbicide resistance, water stress tolerance, and agronomic traits aswell as grain quality traits. Transformation can also be used to insertDNA sequences which control or help control male-sterility. DNAsequences native to wheat as well as non-native DNA sequences can betransformed into wheat and used to modulate levels of native ornon-native proteins. Anti-sense technology, RNA interference, variouspromoters, targeting sequences, enhancing sequences, and other DNAsequences can be inserted into the wheat genome for the purpose ofmodulating the expression of proteins. Exemplary genes implicated inthis regard include, but are not limited to, those categorized below.

1. Genes that confer resistance to pests or disease include one or moreof those described as follows in (A)-(V):

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266: 789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262: 1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinoset al., Cell 78: 1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae); McDowell & Woffenden, (2003) Trends Biotechnol.21(4): 178-83 and Toyoda et al., (2002) Transgenic Res. 11(6):567-82. Aplant resistant to a disease is one that is more resistant to a pathogenas compared to the wild type plant.

Fusarium head blight along with deoxynivalenol both produced by thepathogen Fusarium graminearum Schwabe have caused devastating losses inwheat production. Genes expressing proteins with antifungal action canbe used as transgenes to prevent Fusarium head blight. Various classesof proteins have been identified. Examples include endochitinases,exochitinases, glucanases, thionins, thaumatin-like proteins, osmotins,ribosome inactivating proteins, flavoniods, lactoferricin. Duringinfection with Fusarium graminearum deoxynivalenol (DON) is produced.There is evidence that production of deoxynivalenol increases thevirulence of the disease. Genes with properties for detoxification ofdeoxynivalenol (Adam and Lemmens, In International Congress on MolecularPlant-Microbe Interactions, 1996; McCormick et al. Appl. Environ. Micro.65:5252-5256, 1999) have been engineered for use in wheat. A syntheticpeptide that competes with deoxynivalenol has been identified (Yuan etal., Appl. Environ. Micro. 65:3279-3286, 1999). Changing the ribosomesof the host so that they have reduced affinity for deoxynivalenol hasalso been used to reduce the virulence of the Fusarium graminearum.

Genes used to help reduce Fusarium head blight include but are notlimited to Tri101 (Fusarium), PDR5 (yeast), tlp-1(oat), tlp-2(oat), leaftlp-1 (wheat), tlp (rice), tlp-4 (oat), endochitinase, exochitinase,glucanase (Fusarium), permatin (oat), seed hordothionin (barley),alpha-thionin (wheat), acid glucanase (alfalfa), chitinase (barley andrice), class beta II-1,3-glucanase (barley), PR5/tlp (arabidopsis),zeamatin (maize), type 1 RIP (barley), NPR1 (arabidopsis), lactoferrin(mammal), oxalyl-CoA-decarboxylase (bacterium), IAP (baculovirus), ced-9(C. elegans), and glucanase (rice and barley) (Dahleen, L. S., Okubara,P. A. and A. E. Blechl (2001) Transgenic Approaches to Combat FusariumHead Blight in Wheat and Barley, Crop Science 41:628-637).

(B) A gene conferring resistance to a pest, such as Hessian fly, wheat,stem soft fly, cereal leaf beetle, and/or green bug. For example the H9,H10, and H21 genes.

(C) A gene conferring resistance to disease, including wheat rusts,Septoria tritici, Septoria nodorum, powdery mildew, helminthosporiumdiseases, smuts, bunts, fusarium diseases, bacterial diseases, and viraldiseases.

(D) A Bacillus thuringiensis protein, a derivative thereof or asynthetic polypeptide modeled thereon. See, for example, Geiser et al.,Gene 48: 109 (1986), who disclose the cloning and nucleotide sequence ofa Bt delta-endotoxin gene. Moreover, DNA molecules encodingdelta-endotoxin genes can be purchased from American Type CultureCollection (Rockville, Md.), for example, under ATCC Accession Nos.40098, 67136, 31995 and 31998. Other examples of Bacillus thuringiensistransgenes being genetically engineered are given in the followingpatents and patent applications and hereby are incorporated by referencefor this purpose: U.S. Pat. Nos. 5,188,960, Payne et al, issued Feb. 23,1993; 5,689,052, Brown et al, issued Nov. 18, 1997; 5,880,275, Fischhoffet al., issued Mar. 9, 1999; WO 91/14778, Donovan et al., published Oct.3, 1991; WO 99/31248, English et al., published Jun. 24, 1999; WO01/12731, Bice et al., published Feb. 22, 2001; WO 99/24581, Cardineauet al., published May 20, 1999; WO 97/40162, Narva et al., publishedOct. 30, 1997; and U.S. Pub. Nos. 2002/0151709, Abad et al., publishedOct. 17, 2002; 2003/0177528, Abad et al., published Sep. 18, 2003; and2004/0091500, Ipsen et al., published May 13, 2004.

(E) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344: 458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

(F) An insect-specific peptide which, upon expression, disrupts thephysiology of the affected pest. For example, see the disclosures ofRegan, J. Biol. Chem. 269: 9 (1994) (expression cloning yields DNAcoding for insect diuretic hormone receptor); Pratt et al., Biochem.Biophys. Res. Comm. 163: 1243 (1989) (an allostatin is identified inDiploptera puntata); Chattopadhyay et al. (2004) Critical Reviews inMicrobiology 30 (1): 33-54 2004; Zjawiony (2004) J Nat Prod 67 (2):300-310; Carlini & Grossi-de-Sa (2002) Toxicon, 40 (11):1515-1539; Ussufet al. (2001) Curr Sci. 80 (7): 847-853; and Vasconcelos & Oliveira(2004) Toxicon 44 (4): 385-403. See also U.S. Pat. No. 5,266,317,Tomalski et al., issued Nov. 30, 1993, disclosing genes encodinginsect-specific toxins.

(G) An enzyme responsible for an hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(H) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197, Scott et al., published Feb. 4, 1993 in thename of Scott et al., which discloses the nucleotide sequence of acallase gene. DNA molecules which contain chitinase-encoding sequencescan be obtained, for example, from the ATCC under Accession Nos. 39637and 67152. See also Kramer et al., Insect Biochem. Molec. Biol. 23: 691(1993), who teach the nucleotide sequence of a cDNA encoding tobaccohookworm chitinase, and Kawalleck et al., Plant Molec. Biol. 21: 673(1993), who provide the nucleotide sequence of the parsley ubi4-2polyubiquitin gene, U.S. Pat. Nos. 7,145,060, Muller et al., issued Dec.5, 2006; 7,087,810, Muller et al., issued Aug. 8, 2006; and 6,563,020,Simmons et al., issued May 13, 2003.

(I) A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24: 757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104: 1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

(J) A hydrophobic moment peptide. See WO 95/16776, Putman et al.,published Jun. 22, 1995, and U.S. Pat. No. 5,580,852, Putnam et al.,issued Dec. 3, 1996 (disclosure of peptide derivatives of Tachyplesinwhich inhibit fungal plant pathogens) and WO 95/18855, Rao et al.,published Jul. 13, 1995, and U.S. Pat. No. 5,607,914, Rao et al., issuedMar. 4, 1997 (teaches synthetic antimicrobial peptides that conferdisease resistance).

(K) A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure by Jaynes et al., Plant Sci. 89: 43 (1993),of heterologous expression of a cecropin-beta lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

(L) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. Rev. Phytopathol.28: 451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus.

(M) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al, Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

(N) A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366: 469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

(O) A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo alpha-1,4-D-polygalacturonasesfacilitate fungal colonization and plant nutrient release bysolubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See Lamb etal., Bio/Technology 10: 1436 (1992). The cloning and characterization ofa gene which encodes a bean endopolygalacturonase-inhibiting protein isdescribed by Toubart et al., Plant J. 2: 367 (1992).

(P) A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10: 305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

(Q) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, S., Current Biology,5(2):128-131 (1995), Pieterse & Van Loon (2004) Curr. Opin. Plant Bio.7(4):456-64 and Somssich (2003) Cell 11 3(7):81 5-6.

(R) Antifungal genes (Cornelissen and Melchers, Pl. Physiol.101:709-712, (1993) and Parijs et al., Planta 183:258-264, (1991) andBushnell et al., Can. J. of Plant Path. 20(2):137-149 (1998). Also seeU.S. Pat. No. 6,875,907, Simmons et al, issued Apr. 5, 2005.

(S) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see U.S. Pat. No. 5,792,931, Duvick et al., issued Aug. 11,1998.

(T) Cystatin and cysteine proteinase inhibitors. See U.S. Pat. No.7,205,453, Altier et al., issued Apr. 17, 2007.

(U) Defensin genes. See WO 03/000863, Cahoon et al., published Jan. 3,2003, and U.S. Pat. No. 6,911,577, Simmons et al., issued Jun. 28, 2005.

(V) Genes conferring resistance to nematodes. See WO 03/033651, Hu etal., published Apr. 24, 2003, and Urwin et. al., Planta 204:472-479(1998), Williamson (1999) Curr Opin Plant Bio. 2(4):327-31.

2. Genes that confer resistance to a herbicide include one or more ofthose described as follows in (A)-(E):

(A) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants (see, e.g., Hattori et al. (1995)Mol Gen Genet. 246:419). Other genes that confer tolerance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota et al. (1994)Plant PhysiolPlant Physiol 106:17), genes for glutathione reductase andsuperoxide dismutase (Aono et al. (1995) Plant Cell Physiol 36:1687, andgenes for various phosphotransferases (Datta et al. (1992) Plant MolBiol 20:619).

(B) A herbicide that inhibits the growing point or meristem, such as animidazalinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7: 1241 (1988), and Miki et al., Theor. Appl. Genet. 80: 449(1990), respectively. See also, U.S. Pat. Nos. 5,605,011, Bedbrook etal., issued Feb. 25, 1997; 5,013,659, Bedbrook et al., issued May 7,1991; 5,141,870, Bedbrook et al., issued Aug. 25, 1992; 5,767,361,Dietrich, issued Jun. 16, 1998; 5,731,180, Dietrich, issued Mar. 24,1998; 5,304,732, Anderson et al., issued Apr. 19, 1994; 4,761,373,Anderson et al, issued Aug. 2, 1988; 5,331,107, Anderson et al., issuedJul. 19, 1994; 5,928,937, Kakefuda et al., issued Jul. 27, 1999; and5,378,824, Bedbrook et al., issued Jan. 3, 1995; and internationalpublication WO 96/33270, Kakefuda et al., published Oct. 24, 1996, whichare incorporated herein by reference for this purpose.

(C) Glyphosate (resistance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus phosphinothricin acetyl transferase (bar) genes), andpyridinoxy or phenoxy proprionic acids and cycloshexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835,Shah et al., issued Jul. 10, 1990, which discloses the nucleotidesequence of a form of EPSPS which can confer glyphosate resistance. U.S.Pat. No. 5,627,061, Barry et al., issued May 6, 1997 also describesgenes encoding EPSPS enzymes. See also U.S. Pat. Nos. 6,566,587, Lebrunet al., issued May 20, 2003; 6,338,961, DeRose et al., issued Jan. 15,2002; 6,248,876, Barry et al., issued Jun. 19, 2001; 6,040,497, Spenceret al., issued Mar. 21, 2000; 5,804,425, Barry et al., issued Sep. 8,1998; 5,633,435, Barry et al., issued May 27, 1997; 5,145,783, Kishoreet al., issued Sep. 8, 1992; 4,971,908, Kishore et al., issued Nov. 20,1990; 5,312,910, Kishore et al., issued May 17, 1994; 5,188,642, Shah etal., issued Feb. 23, 1993; 4,940,835, Shah et al., issued Jul. 10, 1990;5,866,775, Eichholtz et al, issued Feb. 2, 1999; 6,225,114, Eichholtz etal, issued May 1, 2001; 6,130,366, Herrera-Estrella et al., issued Oct.10, 2000; 5,310,667, Eichholtz et al., issued May 10, 1994; 4,535,060,Comai, issued Aug. 13, 1985; 4,769,061, Comai, issued Sep. 6, 1988;5,633,448, Lebrun et al., issued May 27, 1997; 5,510,471, Lebrun et al.,issued Apr. 23, 1996; RE 36,449, Lebrun et al., issued Dec. 14, 1999; RE37,287, Lebrun et al., issued Jul. 17, 2001; and U.S. Pat. No.5,491,288, Chaubet et al., issued Feb. 13, 1996; and EP 1173580, Hawkeset al., Jan. 23, 2002; WO 01/66704, Baerson et al., published Sep. 13,2001; EP 1173581 and EP 1173582, Hawkes et al., published Jan. 23, 2002,which are incorporated herein by reference for this purpose. Glyphosateresistance is also imparted to plants that express a gene that encodes aglyphosate oxido-reductase enzyme as described more fully in U.S. Pat.Nos. 5,776,760, Barry et al., issued Jul. 7, 1998 and 5,463,175, Barryet al., issued Oct. 31, 1995, which are incorporated herein by referencefor this purpose. In addition glyphosate resistance can be imparted toplants by the over expression of genes encoding glyphosateN-acetyltransferase. See, for example, U.S. Pat. No. 7,462,481, Castleet al., issued Dec. 9, 2008. A DNA molecule encoding a mutant aroA genecan be obtained under ATCC accession No. 39256, and the nucleotidesequence of the mutant gene is disclosed in U.S. Pat. No. 4,769,061,Comai, issued Sep. 6, 1988. European Publication No. EP 0333033, Kumadaet al., published Sep. 20, 1989, and U.S. Pat. No. 4,975,374, Goodman etal., issued Dec. 4, 1990, disclose nucleotide sequences of glutaminesynthetase genes which confer resistance to herbicides such asL-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in EP 0242246 andEP 0242236, Leemans et al., published Oct. 21, 1987. De Greef et al.,Bio/Technology 7: 61 (1989), describe the production of transgenicplants that express chimeric bar genes coding for phosphinothricinacetyl transferase activity. See also, U.S. Pat. Nos. 5,969,213, Adamset al., issued Oct. 19, 1999; 5,489,520, Adams et al., issued Feb. 6,1996; 5,550,318, Adams et al., issued Aug. 27, 1996; 5,874,265, Adams etal., issued Feb. 23, 1999; 5,919,675, Adams et al., issued Jul. 6, 1999;5,561,236, Leemans et al., issued Oct. 1, 1996; 5,648,477, Leemans etal., issued Jul. 15, 1997; 5,646,024, Leemans et al., issued Jul. 8,1997; 6,177,616, Bartsch et al., issued Jan. 23, 2001; and U.S. Pat. No.5,879,903, Strauch et al., issued Mar. 9, 1999, which are incorporatedherein by reference for this purpose. Exemplary genes conferringresistance to phenoxy proprionic acids and cycloshexones, such assethoxydim and haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3 genesdescribed by Marshall et al., Theor. Appl. Genet. 83: 435 (1992).

(D) An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3: 169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648, Stalker,issued Mar. 7, 1989, and DNA molecules containing these genes areavailable under ATCC Accession Nos. 53435, 67441 and 67442. Cloning andexpression of DNA coding for a glutathione S-transferase is described byHayes et al., Biochem. J. 285:173 (1992).

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306, Ward et al.,issued Sep. 11, 2001; 6,282,837, Ward et al., issued Sep. 4, 2001; and5,767,373; Ward et al., issued Jun. 16, 1998; and WO 01/12825, Johnsonet al., published Feb. 22, 2001.

3. Genes that confer or improve grain quality include one or more ofthose described as follows in (A)-(E):

(A) Altered fatty acids, for example, by (1) Down-regulation ofstearoyl-ACP desaturase to increase stearic acid content of the plant.See Knultzon et al., Proc. Natl. Acad. Sci. USA 89: 2624 (1992) and WO99/64579, Shen, published Dec. 16, 1999 (Genes for Desaturases to AlterLipid Profiles in Corn), (2) Elevating oleic acid via FAD-2 genemodification and/or decreasing linolenic acid via FAD-3 genemodification (see U.S. Pat. Nos. 6,063,947, DeBonte et al., issued May16, 2000; 6,323,392, Charne, issued Nov. 27, 2001; 6,372,965, Lightneret al., issued Apr. 16, 2002, and WO 93/11245, Browse et al., publishedJun. 10, 1993), (3) Altering conjugated linolenic or linoleic acidcontent, such as in WO 01/12800, Cahoon et al., published Feb. 22, 2001,(4) Altering LEC1, AGP, Dek1, Superal1, mi1ps, various lpa genes such aslpa1, lpa3, hpt or hggt. For example, see WO 02/42424, Lappegard,published May 30, 2002; WO 98/22604, Singletary et al., published May28, 1998; WO 03/011015, Tarczynski et al., published Feb. 13, 2003; U.S.Pat. Nos. 6,423,886; Singletary et al., issued Jul. 23, 2002; 6,197,561,Martino-Catt et al., issued Mar. 6, 2001; 6,825,397, Lowe et al., issuedNov. 30, 2004; U.S. Pub. Nos. 2003/0079247, Shi et al., published Apr.24, 2003; 2003/0204870, Allen et al., published Oct. 30, 2003; WO02/057439, Cahoon et al., published Jul. 25, 2002; WO 03/011015,Tarczynski et al., published Feb. 13, 2003; and Rivera-Madrid, R. etal., Proc. Natl. Acad. Sci. 92:5620-5624 (1995).

(B) Altered phosphorus content, for example, by the (1) Introduction ofa phytase-encoding gene would enhance breakdown of phytate, adding morefree phosphate to the transformed plant. For example, see VanHartingsveldt et al., Gene 127: 87 (1993), for a disclosure of thenucleotide sequence of an Aspergillus niger phytase gene. (2)Up-regulation of a gene that reduces phytate content. In maize, this,for example, could be accomplished, by cloning and then re-introducingDNA associated with one or more of the alleles, such as the LPA alleles,identified in maize mutants characterized by low levels of phytic acid,such as in Raboy et al., Maydica 35: 383 (1990) and/or by alteringinositol kinase activity as in WO 02/059324, Shi et al., published Aug.1, 2002; U.S. Pub. No. 2003/0009011, Shi et al., published Jan. 9, 2003;WO 03/027243, Shi et al., published Apr. 3, 2003; U.S. Pub. No.2003/0079247, Shi et al., published Apr. 24, 2003; WO 99/05298,Martino-Catt et al., published Feb. 4, 199; U.S. Pat. Nos. 6,197,561,Martino-Catt et al., issued Mar. 6, 2001; 6,291,224, Martino-Catt etal., issued Sep. 18, 2001; 6,391,348, Stilborn et al., issued May 21,2002; WO 2002/059324, Shi et al., published Aug. 1, 2002; WO 98/45448,Hitz et al., published Oct. 15, 1998; WO 99/55882, Cahoon et al.,published Nov. 4, 1999; and WO 01/04147, Cahoon, published Jan. 18,2001.

(C) Altered carbohydrates effected, for example, by altering a gene foran enzyme that affects the branching pattern of starch or a genealtering thioredoxin (See U.S. Pat. No. 6,531,648, Lanahan et al.,issued Mar. 11, 2003). See Shiroza et al., J. Bacteriol. 170: 810 (1988)(nucleotide sequence of Streptococcus mutans fructosyltransferase gene),Steinmetz et al., Mol. Gen. Genet. 200: 220 (1985) (nucleotide sequenceof Bacillus subtilis levansucrase gene), Pen et al., Bio/Technology 10:292 (1992) (production of transgenic plants that express Bacilluslicheniformis alpha-amylase), Elliot et al., Plant Molec. Biol. 21: 515(1993) (nucleotide sequences of tomato invertase genes), Sogaard et al.,J. Biol. Chem. 268: 22480 (1993) (site-directed mutagenesis of barleyalpha-amylase gene), and Fisher et al., Plant Physiol. 102: 1045 (1993)(maize endosperm starch branching enzyme II), WO 99/10498, Helentjariset al., published Mar. 4, 1999 (improved digestibility and/or starchextraction through modification of UDP-D-xylose 4-epimerase, Fragile 1and 2, Ref1, HCHL, C4H), U.S. Pat. No. 6,232,529, Singletary et al.,issued May 15, 2001, (method of producing high oil seed by modificationof starch levels (AGP)). The fatty acid modification genes mentionedabove may also be used to affect starch content and/or compositionthrough the interrelationship of the starch and oil pathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see U.S. Pat. No. 6,787,683,Penna et al., issued Sep. 7, 2004; U.S. Pub. No. 2004/0034886, Cahoon etal., published Feb. 19, 2004; and WO 00/68393, Della Penna et al.,published Nov. 16, 2000, involving the manipulation of antioxidantlevels through alteration of a phytl prenyl transferase (ppt), WO03/082899, Cahoon et al., published Oct. 9, 2003, through alteration ofa homogentisate geranyl geranyl transferase (hggt).

(E) Altered essential seed amino acids. For example, see U.S. Pat. Nos.6,127,600, Beach et al., issued Oct. 3, 2000 (method of increasingaccumulation of essential amino acids in seeds); 6,080,913, Tarczynskiet al., issued Jun. 27, 2000 (binary methods of increasing accumulationof essential amino acids in seeds); 5,990,389, Rao et al., issued Nov.23, 1999 (high lysine); WO 99/40209, Jung et al., published Aug. 12,1999 (alteration of amino acid compositions in seeds); WO 99/29882, Raoet al., published Jun. 17, 1999 (methods for altering amino acid contentof proteins); U.S. Pat. No. 5,850,016, Jung et al., issued Dec. 15, 1998(alteration of amino acid compositions in seeds); WO 98/20133, Rao etal., published May 14, 1998 (proteins with enhanced levels of essentialamino acids); U.S. Pat. Nos. 5,885,802, Rao, issued Mar. 23, 1999 (highmethionine); 5,885,801, Rao, issued Mar. 23, 1999 (high threonine);6,664,445, Falco et al., issued Dec. 16, 2003 (plant amino acidbiosynthetic enzymes); 6,459,019, Falco, issued Oct. 1, 2002 (increasedlysine and threonine); 6,441,274, Cahoon et al., issued Aug. 27, 2002(plant tryptophan synthase beta subunit); 6,346,403, Rafalski et al.,issued Feb. 12, 2002 (methionine metabolic enzymes); 5,939,599, Chui etal., issued Aug. 17, 1999 (high sulfur); 5,912,414, Falco et al., issuedJun. 15, 1999 (increased methionine); WO 98/56935, Falco et al., Dec.17, 1998 (plant amino acid biosynthetic enzymes); WO 98/45458,Gutteridge, published Oct. 15, 1998 (engineered seed protein havinghigher percentage of essential amino acids); WO 98/42831, Falco et al.,published Oct. 1, 1998 (increased lysine); U.S. Pat. No. 5,633,436,Wandelt, issued May 27, 1997 (increasing sulfur amino acid content);U.S. Pat. No. 5,559,223, Falco et al., issued Sep. 24, 1996 (syntheticstorage proteins with defined structure containing programmable levelsof essential amino acids for improvement of the nutritional value ofplants); WO 96/01905, Falco, published Jan. 25, 1996 (increasedthreonine); WO 95/15392, Falco et al., Jun. 8, 1995 (increased lysine);U.S. Pub. Nos. 2003/0163838, Dhugga et al., published Aug. 28, 2003;2003/0150014, Dhugga et al., published Aug. 7, 2003; 2004/0068767,Dhugga et al., published Apr. 8, 2004; U.S. Pat. No. 6,803,498, Dhuggaet al., issued Oct. 12, 2004; WO 01/79516, Dhugga et al., published Oct.25, 2001; and WO 00/09706, Dhugga et al., published Feb. 24, 2000 (CesA: cellulose synthase); U.S. Pat. Nos. 6,194,638, Dhugga et al., issuedFeb. 27, 2001 (hemicellulose); 6,399,859, Nichols et al., issued Jun. 4,2002; and U.S. Pub. No. 2004/0025203, Singletary et al., published Feb.5, 2004 (UDPGdH).

4. Genes that control male-sterility.

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465, Brar et al., issued Mar. 31, 1987; and 4,727,219, Brar etal., issued Feb. 23, 1988 and chromosomal translocations as described byPatterson in U.S. Pat. Nos. 3,861,079, Patterson, issued Jan. 21, 1975,and 3,710,511, Patterson, issued Jan. 16, 1973. In addition to thesemethods, U.S. Pat. No. 5,432,068, Albertsen et al., issued Jul. 11,1995, describes a system of nuclear male sterility which includesidentifying a gene which is critical to male fertility; silencing thisnative gene which is critical to male fertility; removing the nativepromoter from the essential male fertility gene and replacing it with aninducible promoter; inserting this genetically engineered gene back intothe plant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing, or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

Such genes and methods include one or more of those described as followsin (A)-(C).

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN—Ac-PPT (WO 01/29237, Quandt et al., published Apr. 26, 2001).

(B) Introduction of various stamen-specific promoters (WO 92/13956,Michiels et al., published Aug. 20, 1992; WO 92/13957, DeBeuckeleer etal., published Aug. 20, 1992).

(C) Introduction of the barnase and the barstar gene (Paul et al. PlantMol. Biol. 19:611-622, 1992).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341, Albertsen et al., issued Jan.12, 1999; 6,297,426, Albertsen et al., issued Oct. 2, 2001; 5,478,369,Albertsen et al., issed Dec. 26, 1995; 5,824,524, Albertsen et al.,issued Oct. 20, 1998; 5,850,014, Albertsen et al., issued Dec. 15, 1998;6,265,640, Albertsen et al., issued Jul. 24, 2001; all of which arehereby incorporated by reference.

5. Genes that create a site for site specific DNA integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.For example, see Lyznik, et al., Site-Specific Recombination for GeneticEngineering in Plants, Plant Cell Rep (2003) 21:925-932 and WO 99/25821,Baszczynski et al., published May 27, 1999, which are herebyincorporated by reference. Other systems that may be used include theGin recombinase of phage Mu (Maeser et al., 1991; Vicki Chandler, TheMaize Handbook ch. 118 (Springer-Verlag 1994), the Pin recombinase of E.coli (Enomoto et al., 1983), and the R/RS system of the pSR1 plasmid(Araki et al., 1992).

6. Genes that affect abiotic stress resistance (including but notlimited to flowering, ear and seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress.

For example, see: WO 00/73475, LaPorte et al., published Dec. 7, 2000,where water use efficiency is altered through alteration of malate; U.S.Pat. Nos. 5,892,009, Thomashow et al, issued Apr. 6, 1999; 5,965,705,Thomashow et al., issued Oct. 12, 1999; 5,929,305, Thomashow et al.,issued Jul. 27, 1999; 5,891,859, Thomashow et al., issued Apr. 6, 1999;6,417,428, Thomashow et al., issued Jul. 9, 2002; 6,664,446, Heard etal., issued Dec. 16, 2003; 6,706,866, Thomashow et al., issued Mar. 16,2004; 6,717,034, Jiang, issued Apr. 6, 2004; 6,801,104, Zhu et al.,issued Oct. 5, 2004; WO 2000/060089, Fromm et al., published Oct. 12,2000; WO 2001/026459, Ratcliffe et al., published Apr. 19, 2001; WO2001/035725, Jiang et al., published May 25, 2001; WO 2001/035727,Rueber et al., published May 25, 2001; WO 2001/036444, Riechmann et al.,published May 25, 2001; WO 2001/036597, Creelman et al., published May25, 2001; WO 2001/036598, Pineda et al., published May 25, 2001; WO2002/015675, Pilgrim et al., Feb. 28, 2002; WO 2002/077185, Reuber,published Oct. 3, 2002; WO 2002/079403, Cai-Zhong, published Oct. 10,2002; WO 2003/013227, Ratcliffe et al., published Feb. 20, 2003; WO2003/013228, Heard et al., published Feb. 20, 2003; WO 2003/014327,Reuber et al., published Feb. 20, 2003; WO 2004/031349, Jiang et al.,published Apr. 15, 2004; WO 2004/076638, Sherman et al., published Sep.10, 2004; WO 98/09521, Thomashow et al., published Mar. 12, 1998; and WO99/38977, Stockinger et al., published Aug. 5, 1999, describing genes,including CBF genes and transcription factors effective in mitigatingthe negative effects of freezing, high salinity, and drought on plants,as well as conferring other positive effects on plant phenotype; U.S.Pub. No. 2004/0148654, Helentjaris, published Jul. 29, 2004 and WO01/36596, Helentjaris, published May 25, 2001, where abscisic acid isaltered in plants resulting in improved plant phenotype such asincreased yield and/or increased tolerance to abiotic stress; WO2000/063401, Habben et al., published Oct. 26, 2000; WO 04/090143,Habben et al., published Oct. 21, 2004; U.S. Pub. No. 2004/0237147,Habben et al., published Nov. 25, 2004; and U.S. Pat. No. 6,992,237,Habben et al., issued Jan. 31, 2006, where cytokinin expression ismodified resulting in plants with increased stress tolerance, such asdrought tolerance, and/or increased yield. Also see WO 0202776, Tiam etal., published Jan. 10, 2002; WO 2003/052063, Campbell et al., publishedJun. 26, 2003; JP 2002281975, Hitoshi et al., published Oct. 2, 2002;U.S. Pat. No. 6,084,153, Good et al., issued Jul. 4, 2000; WO 01/64898,Odom et al., published Sep. 7, 2001; U.S. Pat. Nos. 6,177,275, Coruzziet al., issued Jan. 23, 2001 and 6,107,547, Coruzzi et al., issued Aug.22, 2000 (enhancement of nitrogen utilization and altered nitrogenresponsiveness). For ethylene alteration, see U.S. Pub. Nos.2004/0128719, Klee et al., published Jul. 1, 2004; 2003/0166197, Eckeret al., published Sep. 4, 2003; and WO 2000/32761, Ecker et al.,published Jun. 8, 2000. For plant transcription factors ortranscriptional regulators of abiotic stress, see e.g. U.S. Pub. Nos.2004/0098764, Heard et al., published May 20, 2004; or 2004/0078852,Thomashow et al., published, Apr. 22, 2004.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see e.g. WO97/49811, Coupland et al., published Dec. 31, 1997; (LHY); WO 98/56918,Coupland, published Dec. 17, 1998 (ESD4); WO 97/10339, Bradley et al.,published Mar. 20, 1997; and U.S. Pat. Nos. 6,573,430, Bradley et al.,issued Jun. 3, 2003 (TFL); 6,713,663, Weigel et al., issued Mar. 30,20044 (FT); WO 96/14414, Coupland et al., published May 17, 1996 (CON);WO 96/38560, Dean et al., published Dec. 5, 1996; WO 01/21822, Dean etal., published Mar. 29, 2001 (VRN1); WO 00/44918, Dean et al., publishedAug. 3, 2000 (VRN2); WO 99/49064, Coupland et al., published Sep. 30,1999 (GI); WO 00/46358, Johanson et al., published Aug. 10, 2000 (FRI);WO 97/29123, Harberd et al., published Aug. 14, 1997; U.S. Pat. Nos.6,794,560, Harberd et al., issued Sep. 21, 2004; 6,307,126, Harberd etal., issued Oct. 23, 2001 (GAI); WO 99/09174, Harberd et al., publishedFeb. 25, 1999 (D8 and Rht); and WO 2004/076638, Sherman et al.,published Sep. 10, 2004; and WO 2004/031349, Jiang et al., publishedApr. 15, 2004 (transcription factors).

7. Genes that confer agronomic enhancements, nutritional enhancements,or industrial enhancements include one or more of those described asfollows in (A) and (B).

(A) Improved tolerance to water stress from drought or high salt watercondition. The HVA1 protein belongs to the group 3 LEA proteins thatinclude other members such as wheat pMA2005 (Curry et al., 1991; Curryand Walker-Simmons, 1993), cotton D-7 (Baker et al., 1988), carrot Dc3(Seffens et al., 1990), and rape pLEA76 (Harada et al., 1989). Theseproteins are characterized by 11-mer tandem repeats of amino aciddomains which may form a probable amphophilic alpha-helical structurethat presents a hydrophilic surface with a hydrophobic stripe (Baker etal., 1988; Dure et al., 1988; Dure, 1993). The barley HVA1 gene and thewheat pMA2005 gene (Curry et al., 1991; Curry and Walker-Simmons, 1993)are highly similar at both the nucleotide level and predicted amino acidlevel. These two monocot genes are closely related to the cotton D-7gene (Baker et al., 1988) and carrot Dc3 gene (Seffens et al., 1990)with which they share a similar structural gene organization (Straub etal., 1994). There is, therefore, a correlation between LEA geneexpression or LEA protein accumulation with stress tolerance in a numberof plants. For example, in severely dehydrated wheat seedlings, theaccumulation of high levels of group 3 LEA proteins was correlated withtissue dehydration tolerance (Ried and Walker-Simmons, 1993). Studies onseveral Indica varieties of rice showed that the levels of group 2 LEAproteins (also known as dehydrins) and group 3 LEA proteins in rootswere significantly higher in salt-tolerant varieties compared withsensitive varieties (Moons et al., 1995). The barley HVA1 gene wastransformed into wheat. Transformed wheat plants showed increasedtolerance to water stress, (Sivamani, E. et al. Plant Science 2000, V.155 pl-9 and U.S. Pat. No. 5,981,842, Wu et al., issued Nov. 9, 1999.)

(B) Another example of improved water stress tolerance is throughincreased mannitol levels via the bacterial mannitol-1-phosphatedehydrogenase gene. To produce a plant with a genetic basis for copingwith water deficit, Tarczynski et al. (Proc. Natl. Acad. Sci. USA, 89,2600 (1992); WO 92/19731, Tarczynski et al., published Nov. 12, 1992;Science, 259, 508 (1993)) introduced the bacterial mannitol-1-phosphatedehydrogenase gene, mtlD, into tobacco cells via Agrobacterium-mediatedtransformation. Root and leaf tissues from transgenic plants regeneratedfrom these transformed tobacco cells contained up to 100 mM mannitol.Control plants contained no detectable mannitol. To determine whetherthe transgenic tobacco plants exhibited increased tolerance to waterdeficit, Tarczynski et al. compared the growth of transgenic plants tothat of untransformed control plants in the presence of 250 mM NaCl.After 30 days of exposure to 250 mM NaCl, transgenic plants haddecreased weight loss and increased height relative to theiruntransformed counterparts. The authors concluded that the presence ofmannitol in these transformed tobacco plants contributed to waterdeficit tolerance at the cellular level. See also U.S. Pat. No.5,780,709, Adams et al., issued Jul. 14, 1998, and WO 92/19731,Tarczynski et al., published Nov. 12, 1992, which are incorporatedherein by reference for this purpose.

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,Glick, B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton,1993) pages 89-119.

Further embodiments of the present technology are the treatment of E0028with a mutagen and the plant produced by mutagenesis of E0028.Information about mutagens and mutagenizing seeds or pollen arepresented in the IAEA's Manual on Mutation Breeding (IAEA, 1977) otherinformation about mutation breeding in wheat can be found in C. F.Konzak, “Mutations and Mutation Breeding” chapter 7B, of Wheat and WheatImprovement, 2nd edition, Ed. Heyne, 1987.

A further embodiment is a backcross conversion of MSU Line E0028. Abackcross conversion occurs when DNA sequences are introduced throughtraditional (non-transformation) breeding techniques, such asbackcrossing. DNA sequences, whether naturally occurring or transgenes,may be introduced using these traditional breeding techniques. Desiredtraits transferred through this process include, but are not limited tonutritional enhancements, industrial enhancements, disease resistance,insect resistance, herbicide resistance, agronomic enhancements, grainquality enhancement, waxy starch, breeding enhancements, seed productionenhancements, and male sterility. Descriptions of some of thecytoplasmic male sterility genes, nuclear male sterility genes, chemicalhybridizing agents, male fertility restoration genes, and methods ofusing the aforementioned are discussed in “Hybrid Wheat by K. A. Lucken(pp. 444-452 In Wheat and Wheat Improvement, ed. Heyne, 1987). Examplesof genes for other traits include: Leaf rust resistance genes (Lr seriessuch as Lr1, Lr10, Lr21, Lr22, Lr22a, Lr32, Lr37, Lr41, Lr42, and Lr43),Fusarium head blight-resistance genes (QFhs.ndsu-3B and QFhs.ndsu-2A),Powdery Mildew resistance genes (Pm21), common bunt resistance genes(Bt-10), and wheat streak mosaic virus resistance gene (Wsm1), Russianwheat aphid resistance genes (Dn series such as Dn1, Dn2, Dn4, Dn5),Black stem rust resistance genes (Sr38), Yellow rust resistance genes(Yr series such as Yr1, YrSD, Yrsu, Yr17, Yr15, YrH52), Aluminumtolerance genes (Alt(BH)), dwarf genes (Rht), vernalization genes (Vrn),Hessian fly resistance genes (H9, H10, H21, H29), grain color genes(R/r), glyphosate resistance genes (EPSPS), glufosinate genes (bar, pat)and water stress tolerance genes (Hva1, mtlD). The trait of interest istransferred from the donor parent to the recurrent parent, in this case,the wheat plant disclosed herein. Single gene traits may result fromeither the transfer of a dominant allele or a recessive allele.Selection of progeny containing the trait of interest is done by directselection for a trait associated with a dominant allele. Selection ofprogeny for a trait that is transferred via a recessive allele requiresgrowing and selfing the first backcross to determine which plants carrythe recessive alleles. Recessive traits may require additional progenytesting in successive backcross generations to determine the presence ofthe gene of interest.

Another embodiment of this technology is a method of developing abackcross conversion E0028 wheat plant that involves the repeatedbackcrossing to MSU Line E0028. The number of backcrosses made may be 2,3, 4, 5, 6 or greater, and the specific number of backcrosses used willdepend upon the genetics of the donor parent and whether molecularmarkers are utilized in the backcrossing program. See, for example, R.E. Allan, “Wheat” in Principles of Cultivar Development, Fehr, W. R. Ed.(Macmillan Publishing Company, New York, 1987) pages 722-723,incorporated herein by reference. Using backcrossing methods, one ofordinary skill in the art can develop individual plants and populationsof plants that retain at least 70%, 75%, 79%, 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or99% of the genetic profile of MSU Line E0028. The percentage of thegenetics retained in the backcross conversion may be measured by eitherpedigree analysis or through the use of genetic techniques such asmolecular markers or electrophoresis. In pedigree analysis, on average50% of the starting germplasm would be passed to the progeny line afterone cross to another line, 75% after backcrossing once, 87.5% afterbackcrossing twice, and so on. Molecular markers could also be used toconfirm and/or determine the recurrent parent used. The backcrossconversion developed from this method may be similar to E0028. Suchsimilarity may be measured by a side by side phenotypic comparison, withdifferences and similarities determined at a 5% significance level. Anysuch comparison should be made in environmental conditions that accountfor the trait being transferred. For example, herbicide should not beapplied in the phenotypic comparison of herbicide resistant backcrossconversion of E0028 to E0028.

Another embodiment of the technology is an essentially derived varietyof E0028. As determined by the UPOV Convention, essentially derivedvarieties may be obtained for example by the selection of a natural orinduced mutant, or of a somaclonal variant, the selection of a variantindividual from plants of the initial variety, backcrossing, ortransformation by genetic engineering. An essentially derived variety ofE0028 is further defined as one whose production requires the repeateduse of variety E0028 or is predominately derived from variety E0028.International Convention for the Protection of New Varieties of Plants,as amended on Mar. 19, 1991, Chapter V, Article 14, Section 5(c).

This technology is also directed to methods for using MSU Line E0028 inplant breeding. One such embodiment is the method of crossing MSU LineE0028 with another variety of wheat to form a first generationpopulation of F1 plants. The population of first generation F1 plantsproduced by this method is also an embodiment of the present technology.This first generation population of F1 plants will comprise anessentially complete set of the alleles of MSU Line E0028. One ofordinary skill in the art can utilize either breeder books or molecularmethods to identify a particular F1 plant produced using MSU Line E0028,and any such individual plant is also encompassed by this technology.These embodiments also cover use of transgenic or backcross conversionsof MSU Line E0028 to produce first generation F1 plants.

A method of developing a E0028-progeny wheat plant comprising crossingE0028 with a second wheat plant and performing a breeding method is alsoan embodiment of the present technology. A specific method for producinga line derived from MSU Line E0028 is as follows. One of ordinary skillin the art would cross MSU Line E0028 with another variety of wheat,such as an elite variety. The F1 seed derived from this cross would begrown to form a homogeneous population. The F1 seed would contain oneset of the alleles from variety E0028 and one set of the alleles fromthe other wheat variety. The F1 genome would be made-up of 50% varietyE0028 and 50% of the other elite variety. The F1 seed would be grown andallowed to self, thereby forming F2 seed. On average the F2 seed wouldhave derived 50% of its alleles from variety E0028 and 50% from theother wheat variety, but various individual plants from the populationwould have a much greater percentage or much lower percentage of theiralleles derived from E0028. See Wang J. and R. Bernardo, 2000, Crop Sci.40:659-665 and Bernardo, R. and A. L. Kahler, 2001, Theor. Appl. Genet.102:986-992. The F2 seed would be grown and selection of plants would bemade based on visual observation and/or measurement of traits. TheE0028-derived progeny that exhibit one or more of the desiredE0028-derived traits would be selected and each plant would be harvestedseparately. This F3 seed from each plant would be grown in individualrows and allowed to self. Then selected rows or plants from the rowswould be harvested and threshed individually. The selections would againbe based on visual observation and/or measurements for desirable traitsof the plants, such as one or more of the desirable E0028-derivedtraits. The process of growing and selection would be repeated anynumber of times until a homozygous E0028-derived wheat plant isobtained. The homozygous E0028-derived wheat plant would containdesirable traits derived from MSU Line E0028, some of which may not havebeen expressed by the other original wheat variety to which MSU LineE0028 was crossed and some of which may have been expressed by bothwheat varieties but now would be at a level equal to or greater than thelevel expressed in MSU Line E0028. The homozygous E0028-derived wheatplants would have, on average, 50% of their genes derived from MSU LineE0028, but various individual plants from the population would have amuch greater percentage of their alleles derived from E0028. Thebreeding process, of crossing, selfing, and selection may be repeated toproduce another population of E0028-derived wheat plants with, onaverage, 25% of their genes derived from MSU Line E0028, but variousindividual plants from the population would have a much greaterpercentage of their alleles derived from E0028. Another embodiment ofthe present technology is a homozygous E0028-derived wheat plant thathas received E0028-derived traits.

The previous example can be modified in numerous ways, for instanceselection may or may not occur at every selfing generation, selectionmay occur before or after the actual self-pollination process occurs, orindividual selections may be made by harvesting individual spikes,plants, rows or plots at any point during the breeding processdescribed. In addition, double haploid breeding methods may be used atany step in the process. The population of plants produced at each andany generation of selfing is also an embodiment of the presenttechnology, and each such population would consist of plants containingapproximately 50% of its genes from MSU Line E0028, 25% of its genesfrom MSU Line E0028 in the second cycle of crossing, selfing, andselection, 12.5% of its genes from MSU Line E0028 in the third cycle ofcrossing, selfing, and selection, and so on.

Another embodiment of this technology is the method of obtaining ahomozygous E0028-derived wheat plant by crossing MSU Line E0028 withanother variety of wheat and applying double haploid methods to the F1seed or F1 plant or to any generation of E0028-derived wheat obtained bythe selfing of this cross.

Still further, this technology also is directed to methods for producingE0028-derived wheat plants by crossing MSU Line E0028 with a wheat plantand growing the progeny seed, and repeating the crossing or selfingalong with the growing steps with the E0028-derived wheat plant from 1to 2 times, 1 to 3 times, 1 to 4 times, or 1 to 5 times. Thus, any andall methods using MSU Line E0028 in breeding are part of thistechnology, including selfing, pedigree breeding, backcrossing, hybridproduction and crosses to populations. Unique starch profiles, molecularmarker profiles and/or breeding records can be used by those of ordinaryskill in the art to identify the progeny lines or populations derivedfrom these breeding methods.

In addition, this technology also encompasses progeny with the same orgreater yield or test weight of E0028, the same or shorter plant height,and the same or greater resistance to leaf rust, powdery mildew, leafblight, and spindle streak mosaic virus of E0028. The expression ofthese traits may be measured by a side by side phenotypic comparison,with differences and similarities determined at a 5% significance level.Any such comparison should be made in the same environmental conditions.

In conjunction with the present technology, a deposit is made of atleast 2500 seeds of MSU Line E0028 with the American Type CultureCollection (ATCC), Manassas, Va. USA, ATCC Patent Deposit DesignationPTA-10223. The seeds were deposited with the ATCC on Jul. 16, 2009. Thedeposit was tested on Jul. 27, 2009 and on that date, the seeds wereviable. Access to this deposit will be available during the pendency ofthe application to the Commissioner of Patents and Trademarks andpersons determined by the Commissioner to be entitled thereto uponrequest.

Upon issue of claims, the Applicant will make available to the public,pursuant to 37 CFR 1.808, a deposit of at least 2500 seeds of varietyE0028 with the American type Culture Collection (ATCC), 10801 UniversityBoulevard, Manassas, Va. 20110-2209. This deposit of the MSU Line E0028will be maintained in the ATCC depository, which is a public depository,for a period of 30 years, or 5 years after the most recent request, orfor the enforceable life of the patent, whichever is longer, and will bereplaced if it becomes nonviable during that period. Additionally,Applicant has or will satisfy all the requirements of 37 C.F.R.§§1.801-1.809, including providing an indication of the viability of thesample. Applicant has no authority to waive any restrictions imposed bylaw on the transfer of biological material or its transportation incommerce. Applicant does not waive any infringement of their rightsgranted under this patent or under the Plant Variety Protection Act (7USC 2321 et seq.).

MSU Line E0028 is registered and described in the Journal of PlantRegistrations in a publication entitled “Registration of ‘Ambassador’Wheat,” which is incorporated herein by reference.

The embodiments described herein are exemplary and not intended to belimiting in describing the full scope of compositions and methods of thepresent technology. Equivalent changes, modifications and variations ofembodiments, materials, compositions and methods can be made within thescope of the present technology, with substantially similar results.

1-2. (canceled)
 3. A product comprising the wheat plant or the portionof the wheat plant produced by growing the seed of soft winter whitewheat variety designated E0028, representative seed of variety E0028deposited under American Type Culture Collection (ATCC) Patent DepositDesignation PTA-10223, wherein the product is selected from the groupconsisting of grain, flour, baked goods, cereals, pasta, beverages,livestock feed, biofuel, straw, and construction materials. 4-35.(canceled)
 36. The product according to claim 3, selected from the groupconsisting of grain, flour, baked goods, cereals, pasta, and beverages.37. The grain or flour product of claim
 36. 38. The grain of the softwinter white wheat variety designated E0028, representative seed ofvariety E0028 deposited under American Type Culture Collection (ATCC)Patent Deposit Designation PTA-10223.
 39. A product comprising the grainof the soft winter white wheat variety designated E0028, representativeseed of variety E0028 deposited under American Type Culture Collection(ATCC) Patent Deposit Designation PTA-10223.
 40. The product accordingto claim 39, selected from the group consisting of grain, flour, bakedgoods, cereals, pasta, and beverages.